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Auswahl der wissenschaftlichen Literatur zum Thema „CRISPR spacers“

Autor: Grafiati

Veröffentlicht am 18. Mai 2024

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Zeitschriftenartikel zum Thema "CRISPR spacers"

Horvath, Philippe, Dennis A. Romero, Anne-Claire Coûté-Monvoisin, Melissa Richards, Hélène Deveau, Sylvain Moineau, Patrick Boyaval, Christophe Fremaux und Rodolphe Barrangou. „Diversity, Activity, and Evolution of CRISPR Loci in Streptococcus thermophilus“. Journal of Bacteriology 190, Nr. 4 (07.12.2007): 1401–12. http://dx.doi.org/10.1128/jb.01415-07.

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ABSTRACT Clustered regularly interspaced short palindromic repeats (CRISPR) are hypervariable loci widely distributed in prokaryotes that provide acquired immunity against foreign genetic elements. Here, we characterize a novel Streptococcus thermophilus locus, CRISPR3, and experimentally demonstrate its ability to integrate novel spacers in response to bacteriophage. Also, we analyze CRISPR diversity and activity across three distinct CRISPR loci in several S. thermophilus strains. We show that both CRISPR repeats and cas genes are locus specific and functionally coupled. A total of 124 strains were studied, and 109 unique spacer arrangements were observed across the three CRISPR loci. Overall, 3,626 spacers were analyzed, including 2,829 for CRISPR1 (782 unique), 173 for CRISPR2 (16 unique), and 624 for CRISPR3 (154 unique). Sequence analysis of the spacers revealed homology and identity to phage sequences (77%), plasmid sequences (16%), and S. thermophilus chromosomal sequences (7%). Polymorphisms were observed for the CRISPR repeats, CRISPR spacers, cas genes, CRISPR motif, locus architecture, and specific sequence content. Interestingly, CRISPR loci evolved both via polarized addition of novel spacers after exposure to foreign genetic elements and via internal deletion of spacers. We hypothesize that the level of diversity is correlated with relative CRISPR activity and propose that the activity is highest for CRISPR1, followed by CRISPR3, while CRISPR2 may be degenerate. Globally, the dynamic nature of CRISPR loci might prove valuable for typing and comparative analyses of strains and microbial populations. Also, CRISPRs provide critical insights into the relationships between prokaryotes and their environments, notably the coevolution of host and viral genomes.

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Toro, Magaly, Guojie Cao, Wenting Ju, Marc Allard, Rodolphe Barrangou, Shaohua Zhao, Eric Brown und Jianghong Meng. „Association of Clustered Regularly Interspaced Short Palindromic Repeat (CRISPR) Elements with Specific Serotypes and Virulence Potential of Shiga Toxin-Producing Escherichia coli“. Applied and Environmental Microbiology 80, Nr. 4 (13.12.2013): 1411–20. http://dx.doi.org/10.1128/aem.03018-13.

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ABSTRACTShiga toxin-producingEscherichia coli(STEC) strains (n= 194) representing 43 serotypes andE. coliK-12 were examined for clustered regularly interspaced short palindromic repeat (CRISPR) arrays to study genetic relatedness among STEC serotypes. A subset of the strains (n= 81) was further analyzed for subtype I-Ecasand virulence genes to determine a possible association of CRISPR elements with potential virulence. Four types of CRISPR arrays were identified. CRISPR1 and CRISPR2 were present in all strains tested; 1 strain also had both CRISPR3 and CRISPR4, whereas 193 strains displayed a short, combined array, CRISPR3-4. A total of 3,353 spacers were identified, representing 528 distinct spacers. The average length of a spacer was 32 bp. Approximately one-half of the spacers (54%) were unique and found mostly in strains of less common serotypes. Overall, CRISPR spacer contents correlated well with STEC serotypes, and identical arrays were shared between strains with the same H type (O26:H11, O103:H11, and O111:H11). There was no association identified between the presence of subtype I-Ecasand virulence genes, but the total number of spacers had a negative correlation with potential pathogenicity (P< 0.05). Fewer spacers were found in strains that had a greater probability of causing outbreaks and disease than in those with lower virulence potential (P< 0.05). The relationship between the CRISPR-cassystem and potential virulence needs to be determined on a broader scale, and the biological link will need to be established.

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Achigar, Rodrigo, Martina Scarrone, Geneviève M. Rousseau, Cécile Philippe, Felipe Machado, Valentina Duvós, María Pía Campot et al. „Ectopic Spacer Acquisition in Streptococcus thermophilus CRISPR3 Array“. Microorganisms 9, Nr. 3 (01.03.2021): 512. http://dx.doi.org/10.3390/microorganisms9030512.

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Streptococcus thermophilus relies heavily on two type II-A CRISPR-Cas systems, CRISPR1 and CRISPR3, to resist siphophage infections. One hallmark of these systems is the integration of a new spacer at the 5′ end of the CRISPR arrays following phage infection. However, we have previously shown that ectopic acquisition of spacers can occur within the CRISPR1 array. Here, we present evidence of the acquisition of new spacers within the array of CRISPR3 of S. thermophilus. The analysis of randomly selected bacteriophage-insensitive mutants of the strain Uy01 obtained after phage infection, as well as the comparison with other S. thermophilus strains with similar CRISPR3 content, showed that a specific spacer within the array could be responsible for misguiding the adaptation complex. These results also indicate that while the vast majority of new spacers are added at the 5′ end of the CRISPR array, ectopic spacer acquisition is a common feature of both CRISPR1 and CRISPR3 systems in S. thermophilus, and it can still provide phage resistance. Ectopic spacer acquisition also appears to have occurred naturally in some strains of Streptococcus pyogenes, suggesting that it is a general phenomenon, at least in type II-A systems.

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van der Ploeg, Jan R. „Analysis of CRISPR in Streptococcus mutans suggests frequent occurrence of acquired immunity against infection by M102-like bacteriophages“. Microbiology 155, Nr. 6 (01.06.2009): 1966–76. http://dx.doi.org/10.1099/mic.0.027508-0.

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Clustered regularly interspaced short palindromic repeats (CRISPR) consist of highly conserved direct repeats interspersed with variable spacer sequences. They can protect bacteria against invasion by foreign DNA elements. The genome sequence of Streptococcus mutans strain UA159 contains two CRISPR loci, designated CRISPR1 and CRISPR2. The aims of this study were to analyse the organization of CRISPR in further S. mutans strains and to investigate the importance of CRISPR in acquired immunity to M102-like phages. The sequences of CRISPR1 and CRISPR2 arrays were determined for 29 S. mutans strains from different persons. More than half of the CRISPR1 spacers and about 35 % of the CRISPR2 spacers showed sequence similarity with the genome sequence of M102, a virulent siphophage specific for S. mutans. Although only a few spacers matched the phage sequence completely, most of the mismatches had no effect on the amino acid sequences of the phage-encoded proteins. The results suggest that S. mutans is often attacked by M102-like bacteriophages, and that its acquisition of novel phage-derived CRISPR sequences goes along with the presence of S. mutans phages in the environment. Analysis of CRISPR1 of M102-resistant mutants of S. mutans OMZ 381 showed that some of them had acquired novel spacers, and the sequences of all but one of these matched the phage M102 genome sequence. This suggests that the acquisition of the spacers contributed to the resistance against phage infection. However, since not all resistant mutants had new spacers, and since the removal of the CRISPR1 array in one of the mutants and in wild-type strains did not lead to loss of resistance to infection by M102, the acquisition of resistance must be based on further elements as well.

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Serbanescu, M. A., M. Cordova, K. Krastel, R. Flick, N. Beloglazova, A. Latos, A. F. Yakunin, D. B. Senadheera und D. G. Cvitkovitch. „Role of the Streptococcus mutans CRISPR-Cas Systems in Immunity and Cell Physiology“. Journal of Bacteriology 197, Nr. 4 (08.12.2014): 749–61. http://dx.doi.org/10.1128/jb.02333-14.

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CRISPR-Cas systems provide adaptive microbial immunity against invading viruses and plasmids. The cariogenic bacteriumStreptococcus mutansUA159 has two CRISPR-Cas systems: CRISPR1 (type II-A) and CRISPR2 (type I-C) with several spacers from both CRISPR cassettes matching sequences of phage M102 or genomic sequences of otherS. mutans. The deletion of thecasgenes of CRISPR1 (ΔC1S), CRISPR2 (ΔC2E), or both CRISPR1+2 (ΔC1SC2E) or the removal of spacers 2 and 3 (ΔCR1SP13E) inS. mutansUA159 did not affect phage sensitivity when challenged with virulent phage M102. Using plasmid transformation experiments, we demonstrated that the CRISPR1-Cas system inhibits transformation ofS. mutansby the plasmids matching the spacers 2 and 3. Functional analysis of thecasdeletion mutants revealed that in addition to a role in plasmid targeting, both CRISPR systems also contribute to the regulation of bacterial physiology inS. mutans. Compared to wild-type cells, the ΔC1S strain displayed diminished growth under cell membrane and oxidative stress, enhanced growth under low pH, and had reduced survival under heat shock and DNA-damaging conditions, whereas the ΔC2E strain exhibited increased sensitivity to heat shock. Transcriptional analysis revealed that the two-component signal transduction system VicR/K differentially modulates expression ofcasgenes within CRISPR-Cas systems, suggesting that VicR/K might coordinate the expression of two CRISPR-Cas systems. Collectively, we providein vivoevidence that the type II-A CRISPR-Cas system ofS. mutansmay be targeted to manipulate its stress response and to influence the host to control the uptake and dissemination of antibiotic resistance genes.

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Pavlova, Yekaterina S., David Paez-Espino, Andrew Yu Morozov und Ilya S. Belalov. „Searching for fat tails in CRISPR-Cas systems: Data analysis and mathematical modeling“. PLOS Computational Biology 17, Nr. 3 (26.03.2021): e1008841. http://dx.doi.org/10.1371/journal.pcbi.1008841.

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Understanding CRISPR-Cas systems—the adaptive defence mechanism that about half of bacterial species and most of archaea use to neutralise viral attacks—is important for explaining the biodiversity observed in the microbial world as well as for editing animal and plant genomes effectively. The CRISPR-Cas system learns from previous viral infections and integrates small pieces from phage genomes called spacers into the microbial genome. The resulting library of spacers collected in CRISPR arrays is then compared with the DNA of potential invaders. One of the most intriguing and least well understood questions about CRISPR-Cas systems is the distribution of spacers across the microbial population. Here, using empirical data, we show that the global distribution of spacer numbers in CRISPR arrays across multiple biomes worldwide typically exhibits scale-invariant power law behaviour, and the standard deviation is greater than the sample mean. We develop a mathematical model of spacer loss and acquisition dynamics which fits observed data from almost four thousand metagenomes well. In analogy to the classical ‘rich-get-richer’ mechanism of power law emergence, the rate of spacer acquisition is proportional to the CRISPR array size, which allows a small proportion of CRISPRs within the population to possess a significant number of spacers. Our study provides an alternative explanation for the rarity of all-resistant super microbes in nature and why proliferation of phages can be highly successful despite the effectiveness of CRISPR-Cas systems.

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Deveau, Hélène, Rodolphe Barrangou, Josiane E. Garneau, Jessica Labonté, Christophe Fremaux, Patrick Boyaval, Dennis A. Romero, Philippe Horvath und Sylvain Moineau. „Phage Response to CRISPR-Encoded Resistance in Streptococcus thermophilus“. Journal of Bacteriology 190, Nr. 4 (07.12.2007): 1390–400. http://dx.doi.org/10.1128/jb.01412-07.

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ABSTRACT Clustered regularly interspaced short palindromic repeats (CRISPR) and their associated genes are linked to a mechanism of acquired resistance against bacteriophages. Bacteria can integrate short stretches of phage-derived sequences (spacers) within CRISPR loci to become phage resistant. In this study, we further characterized the efficiency of CRISPR1 as a phage resistance mechanism in Streptococcus thermophilus. First, we show that CRISPR1 is distinct from previously known phage defense systems and is effective against the two main groups of S. thermophilus phages. Analyses of 30 bacteriophage-insensitive mutants of S. thermophilus indicate that the addition of one new spacer in CRISPR1 is the most frequent outcome of a phage challenge and that the iterative addition of spacers increases the overall phage resistance of the host. The added new spacers have a size of between 29 to 31 nucleotides, with 30 being by far the most frequent. Comparative analysis of 39 newly acquired spacers with the complete genomic sequences of the wild-type phages 2972, 858, and DT1 demonstrated that the newly added spacer must be identical to a region (named proto-spacer) in the phage genome to confer a phage resistance phenotype. Moreover, we found a CRISPR1-specific sequence (NNAGAAW) located downstream of the proto-spacer region that is important for the phage resistance phenotype. Finally, we show through the analyses of 20 mutant phages that virulent phages are rapidly evolving through single nucleotide mutations as well as deletions, in response to CRISPR1.

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Bolotin, Alexander, Benoit Quinquis, Alexei Sorokin und S. Dusko Ehrlich. „Clustered regularly interspaced short palindrome repeats (CRISPRs) have spacers of extrachromosomal origin“. Microbiology 151, Nr. 8 (01.08.2005): 2551–61. http://dx.doi.org/10.1099/mic.0.28048-0.

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Numerous prokaryote genomes contain structures known as clustered regularly interspaced short palindromic repeats (CRISPRs), composed of 25–50 bp repeats separated by unique sequence spacers of similar length. CRISPR structures are found in the vicinity of four genes named cas1 to cas4. In silico analysis revealed another cluster of three genes associated with CRISPR structures in many bacterial species, named here as cas1B, cas5 and cas6, and also revealed a certain number of spacers that have homology with extant genes, most frequently derived from phages, but also derived from other extrachromosomal elements. Sequence analysis of CRISPR structures from 24 strains of Streptococcus thermophilus and Streptococcus vestibularis confirmed the homology of spacers with extrachromosomal elements. Phage sensitivity of S. thermophilus strains appears to be correlated with the number of spacers in the CRISPR locus the strain carries. The authors suggest that the spacer elements are the traces of past invasions by extrachromosomal elements, and hypothesize that they provide the cell immunity against phage infection, and more generally foreign DNA expression, by coding an anti-sense RNA. The presence of gene fragments in CRISPR structures and the nuclease motifs in cas genes of both cluster types suggests that CRISPR formation involves a DNA degradation step.

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Heussler, Gary E., Jon L. Miller, Courtney E. Price, Alan J. Collins und George A. O'Toole. „Requirements for Pseudomonas aeruginosa Type I-F CRISPR-Cas Adaptation Determined Using a Biofilm Enrichment Assay“. Journal of Bacteriology 198, Nr. 22 (29.08.2016): 3080–90. http://dx.doi.org/10.1128/jb.00458-16.

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ABSTRACTCRISPR (clustered regularly interspaced short palindromic repeat)-Cas (CRISPR-associated protein) systems are diverse and found in many archaea and bacteria. These systems have mainly been characterized as adaptive immune systems able to protect against invading mobile genetic elements, including viruses. The first step in this protection is acquisition of spacer sequences from the invader DNA and incorporation of those sequences into the CRISPR array, termed CRISPR adaptation. Progress in understanding the mechanisms and requirements of CRISPR adaptation has largely been accomplished using overexpression ofcasgenes or plasmid loss assays; little work has focused on endogenous CRISPR-acquired immunity from viral predation. Here, we developed a new biofilm-based assay system to enrich forPseudomonas aeruginosastrains with new spacer acquisition. We used this assay to demonstrate thatP. aeruginosarapidly acquires spacers protective against DMS3vir, an engineered lytic variant of the Mu-like bacteriophage DMS3, through primed CRISPR adaptation from spacers present in the native CRISPR2 array. We found that for theP. aeruginosatype I-F system, thecas1gene is required for CRISPR adaptation,recGcontributes to (but is not required for) primed CRISPR adaptation,recDis dispensable for primed CRISPR adaptation, and finally, the ability of a putative priming spacer to prime can vary considerably depending on the specific sequences of the spacer.IMPORTANCEOur understanding of CRISPR adaptation has expanded largely through experiments in type I CRISPR systems using plasmid loss assays, mutants ofEscherichia coli, orcas1-cas2overexpression systems, but there has been little focus on studying the adaptation of endogenous systems protecting against a lytic bacteriophage. Here we describe a biofilm system that allowsP. aeruginosato rapidly gain spacers protective against a lytic bacteriophage. This approach has allowed us to probe the requirements for CRISPR adaptation in the endogenous type I-F system ofP. aeruginosa. Our data suggest that CRISPR-acquired immunity in a biofilm may be one reason that manyP. aeruginosastrains maintain a CRISPR-Cas system.

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Lopatina, Anna, Sofia Medvedeva, Daria Artamonova, Matvey Kolesnik, Vasily Sitnik, Yaroslav Ispolatov und Konstantin Severinov. „Natural diversity of CRISPR spacers of Thermus : evidence of local spacer acquisition and global spacer exchange“. Philosophical Transactions of the Royal Society B: Biological Sciences 374, Nr. 1772 (25.03.2019): 20180092. http://dx.doi.org/10.1098/rstb.2018.0092.

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We investigated the diversity of CRISPR spacers of Thermus communities from two locations in Italy, two in Chile and one location in Russia. Among the five sampling sites, a total of more than 7200 unique spacers belonging to different CRISPR-Cas systems types and subtypes were identified. Most of these spacers are not found in CRISPR arrays of sequenced Thermus strains. Comparison of spacer sets revealed that samples within the same area (separated by few to hundreds of metres) have similar spacer sets, which appear to be largely stable at least over the course of several years. While at further distances (hundreds of kilometres and more) the similarity of spacer sets is decreased, there are still multiple common spacers in Thermus communities from different continents. The common spacers can be reconstructed in identical or similar CRISPR arrays, excluding their independent appearance and suggesting an extensive migration of thermophilic bacteria over long distances. Several new Thermus phages were isolated in the sampling sites. Mapping of spacers to bacteriophage sequences revealed examples of local acquisition of spacers from some phages and distinct patterns of targeting of phage genomes by different CRISPR-Cas systems. This article is part of a discussion meeting issue ‘The ecology and evolution of prokaryotic CRISPR-Cas adaptive immune systems’.

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Mehr Quellen

Dissertationen zum Thema "CRISPR spacers"

Medvedeva, Sofia. „Natural Diversity of CRISPR Spacers“. Electronic Thesis or Diss., Sorbonne université, 2019. http://www.theses.fr/2019SORUS538.

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Le système CRISPR-Cas est un système immunitaire procaryote de type interférence ARN dirigé contre des éléments génétiques mobiles, tels que les virus et les plasmides. Le système consiste en un ou plusieurs loci CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats ; courtes répétitions palindromiques groupées et régulièrement espacées) associés à des protéines Cas (CRISPR-associated proteins) dont ils sont séparés par une séquence dite leader. Toutes les protéines Cas peuvent être fonctionnellement attribuées à des modules d'adaptation, d'expression et d'interférence. L’analyse des spacers CRISPR est une précieuse source d’informations sur les interactions virus-hôte, puisqu’ils correspondent à de courts fragments d’ADN de virus précédemment rencontrés et « enregistrés » dans les loci CRISPR. La comparaison des spacers environnementaux les uns avec les autres et avec des spacers de bases de données ainsi que des séquences de virus nous a permis de tirer plusieurs conclusions : 1) Une dynamique à long terme des spacers CRISPR I-E de E. coli a été étudiée en comparant la diversité des spacers dans les génomes publiés d'E. coli avec des spacers amplifiés à partir du contenu intestinal de mammouth. 2) Les spacers du CRISPRome des communautés naturelles de Thermus, Sulfolobus et Flavobacteries ciblent de préférence des virus isolés de la même source, avec différents systèmes CRISPR-Cas ciblant différents virus. 3) Les données CRISPRome de Flavobacterium et Sulfolobus (Chapitres II et IV) montrent un schéma phylogéographique
CRISPR-Cas is a prokaryotic immunity system against mobile genetic elements, such as viruses and plasmids. The system consists of two components: the clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated proteins (Cas). In the CRISPR array, short fragments of foreign DNA, called spacers, are interleaved with palindromic repeats. During the adaptation stage of the CRISPR-Cas immunity, new spacers are inserted into the CRISPR array, whereas during the expression and interference stages, spacers are transcribed, processed and complexed with Cas proteins to target the complementary foreign DNA or RNA molecules for degradation. Analysis of CRISPR spacers is a valuable source of information about virus-host interactions, particularly powerful when applied to metagenomic data. In this work, we explored the diversity of CRISPR spacers in different natural prokaryotic communities, including extinct Escherichia coli community from a mammoth intestine, Flavobacterium communities from Antarctic surface snow, Thermus communities from four distant hot springs in Italy and Chile, and Sulfolobales community from a Japanese thermal field. The comparison of obtained environmental spacer sets with each other and with spacers from public databases as well as with sequences of viruses allowed us to reach several non-trivial conclusions and to gain insights into virus-host and virus-virus interactions in natural microbial communities

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Vyhovskyi, Danylo. „In vivo studies of CRISPR adaptation mechanism and specificity“. Electronic Thesis or Diss., Sorbonne université, 2023. http://www.theses.fr/2023SORUS729.

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Cette thèse examine les mécanismes de l'immunité adaptative CRISPR-Cas chez les procaryotes, en utilisant principalement le système de type I-E chez Escherichia coli, en se concentrant sur le processus d'acquisition de spacers et la spécificité du système. Elle éclaire la dynamique de génération de spacers et leur intégration dans les CRISPR-arrays, en comparant les modes d'adaptation naïve et primed. L'étude révèle qu'une séquence proximale à un PAM (protospacer adjacent motif) particulier entrave l'acquisition de spacers en mode primed, fournissant un identifiant distinct pour les spacers acquis naturellement. L'étude révèle en outre le rôle des enzymes non-Cas, liées aux voies de réparation de l'ADN, dans la génération et le traitement des spacers, contribuant à l'adaptation et à l'interférence du CRISPR.Un autre volet de l'étude identifie les dangers potentiels posés par les effets hors cible causés par l'enzyme guidée par l'ARN Cas9 (dCas9) qui peut inadvertamment réduire au silence les gènes. Cela se produit lorsqu'il y a une correspondance minimale de seulement quatre nucléotides entre l'ARNg et la cible à l'intérieur de la séquence proximale du PAM, soulignant la nécessité d'une conception expérimentale soigneuse dans la recherche CRISPR-Cas. Dans l'ensemble, la thèse élargit la compréhension des mécanismes moléculaires complexes derrière l'adaptation du CRISPR, soulignant le rôle des protéines non-Cas et l'importance d'un contexte génétique spécifique des séquences proximales à un PAM, conduisant au développement d'outils de génie génétique plus précis et efficaces
This thesis investigates the mechanisms of the CRISPR-Cas adaptive immunity in prokaryotes, primarily using the type I-E system in Escherichia coli, focusing on the spacer acquisition process and the system's specificity. It sheds light on the dynamics of spacers generation and integration into CRISPR arrays, comparing naive and primed adaptation modes. The study reveals that a particular PAM (protospacer adjacent motif) - proximal sequence impedes spacer acquisition in the primed mode, providing a distinct identifier for naturally acquired spacers. The study further reveals the role of non-Cas enzymes, linked to DNA repair pathways, in spacer generation and processing, contributing to CRISPR adaptation and interference.Another course of the study identifies potential hazards posed by off-target effects caused by Cas9 (dCas9) RNA-guided enzyme that can inadvertently silence genes. This occurs when there's a minimal match of just four nucleotides between the gRNA and the target within the PAM-proximal sequence, emphasizing the need for careful experimental design in CRISPR-Cas research.Overall, the thesis expands understanding of the complex molecular mechanisms behind CRISPR adaptation, highlighting the role of non-Cas proteins and the significance of a specific genetic context of seed sequences, leading to the development of more precise and efficient genetic engineering tools

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Jacot-Guillarmod, Paul. „Sobriety of crisp and fuzzy topological spaces“. Thesis, Rhodes University, 2004. http://hdl.handle.net/10962/d1005228.

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The objective of this thesis is a survey of crisp and fuzzy sober topological spaces. We begin by examining sobriety of crisp topological spaces. We then extend this to the L- topological case and obtain analogous results and characterizations to those of the crisp case. We then brie y examine semi-sobriety of (L;M)-topological spaces.

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Stens, Cassandra, Isabella Enoksson und Sara Berggren. „The CRISPR-Cas system“. Thesis, Linköpings universitet, Institutionen för fysik, kemi och biologi, 2020. http://urn.kb.se/resolve?urn=urn:nbn:se:liu:diva-171997.

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Derived from and inspired by the adaptive immune system of bacteria, CRISPR has gone from basic biology knowledge to a revolutionizing biotechnological tool, applicable in many research areas such as medicine, industry and agriculture. The full mechanism of CRISPR-Cas9 was first published in 2012 and various CRISPR-Cas systems have already passed the first stages of clinical trials as new gene therapies. The immense research has resulted in continuously growing knowledge of CRISPR systems and the technique seems to have the potential to greatly impact all life on our planet. Therefore, this literature study aims to thoroughly describe the CRISPR-Cas system, and further suggest an undergraduate laboratory exercise involving gene editing with the CRISPR-Cas9 tool. In this paper, we describe the fundamental technical background of the CRISPR-Cas system, especially emphasizing the most studied CRISPR-Cas9 system, its development and applications areas, as well as highlighting its current limitations and ethical concerns. The history of genetic engineering and the discovery of the CRISPR system is also described, along with a comparison with other established gene editing techniques. This study concludes that a deeper knowledge about CRISPR is important and required since the technique is applicable in many research areas. A laboratory exercise will not only inspire but also provide extended theoretical and practical knowledge for undergraduate students.

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Wong, Shi Pey. „Analysis of the adaptation mechanism in the type II-A CRISPR-Cas system“. Doctoral thesis, Humboldt-Universität zu Berlin, 2019. http://dx.doi.org/10.18452/19806.

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Das RNA-guided adaptive Immunsystem CRISPR (clustered regularly interspaced short palindromic repeats)-Cas (CRISPR-associated) immunisiert prokaryotische Zellen gegenüber mobilen genetischen Elementen (MGEs). Bei der Adaption wird eine kurze Nukleinsäurensequenz (prespacer) von den MGEs gewonnen, verarbeitet und schließlich als spacer in das CRISPR-Array integriert. Cas1 und Cas2, die Hauptbestandteile der Adaption, bilden einen Integrase-Komplex, welcher neue spacer in das CRISPR-Array integriert. Der molekulare Mechanismus für die Adaptiondes Typ II-A Systems, welches cas9, cas1, cas2, csn2 und tracrRNA codiert, ist bis heute nicht vollständig verstanden. Daher untersuchten wir die Anforderungen der verschiedenen Cas-Proteine für den Adaptionsprozess. Wir verifizierten die Adaptions-Aktivität von Typ II-A Systemen des Streptococcus thermophilus LMD-9 anhand von Adaptionsstudien nach Phagen-Infektion. Dabei beobachteten wir höhere Akquisitionsraten im CRISPR3-Lokus im Vergleich zum CRISPR1-Lokus. Unsere Plasmid-basierte Adaptionsstudie bestätigte die Notwendigkeit von Cas9, zusätzlich zu Cas1, Cas2 und Csn2 bei der Adaption. Der yeast two-hybrid und der pull-down Ansatz zeigten sowohl spezifische Interaktionen zwischen den Cas-Proteinen, als auch Interaktionen zwischen Cas-Proteinen sowie DNA-Reparatur Proteinen. Die Regionen der Cas1 und Cas9 Interaktion wurden durch SPOT peptide assay identifiziert. Zusammenfassend weist unsere Studie darauf hin, dass Cas-Proteine sowohl mit Proteinen innerhalb, als auch außerhalb des CRISPR-Cas Systems interagieren, und bietet somit eine Basis für die Erforschung der möglichen Funktionen von DNA-Reparatur Proteinen in CRISPR-Cas Systemen und vice versa.
The RNA guided adaptive immune system CRISPR (clustered regularly interspaced short palindromic repeats) Cas (CRISPR-associated) immunizes prokaryotic cells against mobile genetic elements (MGEs). During spacer acquisition stage, a short nucleic acid sequence (prespacer) is acquired from the MGEs, processed and finally integrated into the CRISPR array as a spacer, which serves as genetic memory to defend against the invasion of the cognate MGEs. The molecular mechanism for the spacer acquisition of the type II A systems, which encode cas9, cas1, cas2, csn2 and tracrRNA, is still not fully understood. Therefore, we investigated the requirement of the different Cas proteins for spacer acquisition. We verified the acquisition activity of the type II A systems of Streptococcus thermophilus LMD 9 via spacer acquisition studies by phage challenge. We observed higher acquisition rates in the CRISPR3 locus compared to the CRISPR1 locus. Our plasmid-based spacer acquisition study confirmed in addition to Cas1, Cas2 and Csn2 the requirement of Cas9 for spacer acquisition. Yeast two hybrid and pull down approaches revealed specific interactions among the Cas proteins, as well as interactions between Cas and DNA repair proteins. The interaction regions of Cas1 with Cas9 were identified by SPOT peptide assay. Altogether, our study suggests that Cas proteins interact with proteins within and beyond the CRISPR Cas systems, and it provides a basis for the investigation of the potential roles of DNA repair proteins in the CRISPR Cas systems and/or vice versa.

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Hille, Frank [Verfasser]. „Investigation of Spacer Acquisition Mechanisms in Type V-A CRISPR-Cas Systems / Frank Hille“. Berlin : Humboldt-Universität zu Berlin, 2020. http://d-nb.info/1215570341/34.

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Ribeiro, Mateus Cotta. „A reprodução crítica do espaço na porção meridional da Serra do Espinhaço de Minas Gerais: modernização do espaço e crise da sociedade do trabalho“. Universidade de São Paulo, 2015. http://www.teses.usp.br/teses/disponiveis/8/8136/tde-15012016-135253/.

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Este estudo é sobre as transformações vividas pelos indivíduos na porção meridional da Serra do Espinhaço do estado de Minas Gerais, Brasil. Estas transformações se manifestam aparentemente como uma expansão da indústria do turismo. Porém, na realidade elas são expressão das determinações impostas ali pela crise da sociedade do trabalho.
This study deals with the social changes experienced by people from the southern area of Serra do Espinhaço, Minas Gerais, Brazil. These changes are related to the increase of the tourism industry however they are determined by crise o work society reproduction.

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Cruz, Murilo Medici Navarro da. „Estado, espaço e acumulação na crise contemporânea“. Universidade de São Paulo, 2006. http://www.teses.usp.br/teses/disponiveis/8/8136/tde-20062007-153251/.

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Este trabalho parte de uma questão posta pela realidade atual: a crise econômica mundial, percebida principalmente no que vem se denominando de financeirização da economia e, conjuntamente a esta crise, aquela do Estado nacional, ambas vistas na perspetiva mundial e, igualmente, na brasileira. Não há, no entanto, o objetivo aqui de se fazer um tratamento histórico ou puramente econômico do problema. Ao invés disso, o que se verá está mais próximo de um exercício teórico e analítico. Este se constitui, em primeiro lugar, de uma discussão aprofundada sobre os fundamentos da economia capitalista que, segundo se entende, são também os fundamentos da crise econômica atual. Em segundo lugar, para se refletir tanto sobre a crise econômica como sobre a política fazse uma reflexão sobre a produção do espaço e o papel do Estado no conjunto da sociedade capitalista. Em terceiro lugar, a conjunção entre o Estado e o capital se mostra identificável na análise do território nacional e seus fundamentos. Por último, o problema da crise atual é posto por meio de uma crítica à idéia de desterritorialização, crítica esta que leva à formulação da hipótese de uma territorialização abstrata, que seria característica da própria crise.
This work starts from a matter put by the current reality: The world economic crisis, specially seem on what has been denominated \"money predomination economy\" and, along with the crisis, the one of national State, both seen under the world\'s perspective and, equally, the Brazilian perspective. Nevertheless, this work does not intend, herein, to give a historical or purely economic treatment of the problem. Instead, what you will see is closer to an analytical and theoretical exercise. This work constitutes, firstly, of a deep discussion about the basis of capital economy which, according to what is understood, also are the basis of the current economic crisis. Secondly, to reflect about the economic crisis as much as politics, is necessary to reflect about the production of space and the State\'s role within the capitalist society. And thirdly, the union between the State and the capital shows itself identifiable on the national territory analysis along with your basis. Last but not least, the current crisis problem is put through a criticism to the lack of focusing the capital on a specific market, criticism that takes us to formulate the hypothesis of an abstract investment of capital, which would be a characteristic of the crisis itself.

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Philipp, Carolin [Verfasser]. „Spaces of Resistance – Framing Crisis and Activism in Athens / Carolin Philipp“. Kassel : Universitätsbibliothek Kassel, 2018. http://d-nb.info/1155948718/34.

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Kreston, Nicholas Alexander. „Post-Keynesian financial spaces, places, and flows : geographies of finance and financial crisis“. Thesis, University of Oxford, 2014. http://ora.ox.ac.uk/objects/uuid:3ea77af2-650c-456a-a4c2-5ee67c83d293.

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This dissertation is a reaction to the public policy failures that culminated in, prolonged, and exacerbated the 2008 financial crisis in the United States. Between the winter of 2007 and the summer of 2009, the US private economy contracted severely. As of the summer of 2014, after a five-year recovery period, employment losses have been restored, but employment growth has not returned the US to its pre-crisis trend. This outcome is not the effect of a transient deviation that regularly happens as the economy moves through the business cycle; nor should the troubles in the US financial sector appear historically anomalous. The world's premier capitalist economy is prone to bouts of financial dysfunction. This feature is not simply a matter of the irrational exuberance of its investors, the euphoria of its speculators, or the folly of its bankers. I argue here that political-economic choices structure the distribution of financial crises at multiple scales. Broadly speaking, I find that the effects of financial crises on growth are uneven, affected by institutional structure, and carry important ramifications for the direction of change in the provision of financial services and its regulatory system. The thesis features four empirical chapters. In the first, I perform an econometric analysis of the effects of a wide variety of financial crises on employment growth by economic sector, for a sample of countries over a thirty-year period. The final three chapters are a case study of the US experience with the 2008 banking crisis and asset market crash, focusing on the role of the banking regulatory system in allocating losses over territory, on the economic performance of metropolitan areas, and on the distribution of losses within the financial sector in two major financial centers, Los Angeles and San Francisco. I reach my conclusions by using standard methodological tools within the sub-field of economic-geography and conceptual insights from the sub-field of financial-geography.

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Bücher zum Thema "CRISPR spacers"

Martin, David. Crisis in space. New York: Ballantine Books, 1986.

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Corporation, British Broadcasting, Hrsg. Crisis in space. London: Severn House by arrangement with the British Broadcasting Corporation, 1986.

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Holt, Michael. Crisis in space. New York: Ballantine Books, 1986.

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ill, Kinsella Pat, Hrsg. Crisis. Minneapolis, MN: Magic Wagon, 2017.

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Nansen, Ralph. Energy crisis: Solution from space. Burlington, Ont: Apogee Books, 2009.

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Reis, Ronald A. Apollo 13: Crisis in space. New York: Chelsea House, 2012.

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Vitalidad y crisis en los espacios públicos de Sevilla. [Seville]: Diputación de Sevilla, 2006.

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Cabannes, Yves, Mike Douglass und Rita Padawangi, Hrsg. Cities in Asia by and for the People. NL Amsterdam: Amsterdam University Press, 2018. http://dx.doi.org/10.5117/9789462985223.

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This book examines the active role of urban citizens in constructing alternative urban spaces as tangible resistance towards capitalist production of urban spaces that continue to encroach various neighborhoods, lanes, commons, public land and other spaces of community life and livelihoods. The collection of narratives presented here brings together research from ten different Asian cities and re-theorises the city from the perspective of ordinary people facing moments of crisis, contestations, and cooperative quests to create alternative spaces to those being produced under prevailing urban processes. The chapters accent the exercise of human agency through daily practices in the production of urban space and the intention is not one of creating a romantic or utopian vision of what a city "by and for the people" ought to be. Rather, it is to place people in the centre as mediators of city-making with discontents about current conditions and desires for a better life.

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Making Humanitarian Spaces Global: Coordinating Crisis Response through the Cluster Approach. [New York, N.Y.?]: [publisher not identified], 2012.

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Bouma, Geiske. Ruimtelijke ordening in crisis: Bijdragen aan de PlanDag 2010. Delft]: [Stichting Planologische Discussiedagen], 2010.

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Buchteile zum Thema "CRISPR spacers"

Davison, Michelle, und Devaki Bhaya. „Creation and Analysis of a Virome: Using CRISPR Spacers“. In Methods in Molecular Biology, 307–16. New York, NY: Springer New York, 2015. http://dx.doi.org/10.1007/978-1-4939-2687-9_20.

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Dupuis, Marie-Ève, Rodolphe Barrangou und Sylvain Moineau. „Procedures for Generating CRISPR Mutants with Novel Spacers Acquired from Viruses or Plasmids“. In Methods in Molecular Biology, 195–222. New York, NY: Springer New York, 2015. http://dx.doi.org/10.1007/978-1-4939-2687-9_13.

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Pallarés, Ana Morcillo. „Crisis and opportunity“. In Manhattan's Public Spaces, 61–69. London: Routledge, 2021. http://dx.doi.org/10.4324/9781003198543-4.

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Mokrousov, Igor, und Nalin Rastogi. „Spacer-Based Macroarrays for CRISPR Genotyping“. In Methods in Molecular Biology, 111–31. New York, NY: Springer New York, 2015. http://dx.doi.org/10.1007/978-1-4939-2687-9_7.

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Burman, Erica. „Lockdown Vistas: Time, Space, Solidarity, Action“. In The Psychology of Global Crises and Crisis Politics, 189–212. Cham: Springer International Publishing, 2021. http://dx.doi.org/10.1007/978-3-030-76939-0_9.

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Jones, Martin, und Kevin Ward. „Excavating the Logic of British Urban Policy: Neoliberalism as the “Crisis of Crisis-Management”“. In Spaces of Neoliberalism, 126–47. Chichester, UK: John Wiley & Sons, Ltd, 2012. http://dx.doi.org/10.1002/9781444397499.ch6.

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Pallarés, Ana Morcillo. „Identity crisis: Lincoln Center and the High Line“. In Manhattan's Public Spaces, 144–69. London: Routledge, 2021. http://dx.doi.org/10.4324/9781003198543-9.

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Martín-Moruno, Dolores. „Crisis? What Crisis? Making Humanitarian Crises Visible in the History of Emotions“. In Making Humanitarian Crises, 1–28. Cham: Springer International Publishing, 2022. http://dx.doi.org/10.1007/978-3-031-00824-5_1.

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AbstractThis introductory chapter carves out a space for the histories of emotions and the senses within humanitarian visual culture, by examining the performativity of images in the construction of humanitarian crises. To this end, it analyses how illustrated pamphlets, cinema talks, photographs, documentary films, graphic novels and virtual reality environments have mobilised the affective responses of audiences, thus creating transnational networks of solidarity from the nineteenth century to present-day society. Furthermore, it questions presentist conceptions of pain, compassion, sympathy and empathy, as well as that these emotions have been the natural reaction of spectators regarding the pain of others. The chapter concludes by showing the potential of investigating related conceptions of indignation, shame, rage and horror in order to advocate for a history of humanitarian experiences.

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Gouw, Arvin M. „CRISPR Challenges and Opportunities for Space Travel“. In Human Enhancements for Space Missions, 19–34. Cham: Springer International Publishing, 2020. http://dx.doi.org/10.1007/978-3-030-42036-9_2.

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Maciocco, Giovanni, und Silvano Tagliagambe. „The Aesthetic Crisis as the Crisis of the Glance“. In People and Space, 13–18. Dordrecht: Springer Netherlands, 2009. http://dx.doi.org/10.1007/978-1-4020-9879-6_3.

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Konferenzberichte zum Thema "CRISPR spacers"

Timkin, P. D., und A. A. Penzin. „An experimental approach for diagnosing cercosporosis using RPA+CRISPR/Cas12a“. In II All-Russian (national) scientific conference with international participation "Russian Science, Innovation, Education". Krasnoyarsk Science and Technology City Hall, 2023. http://dx.doi.org/10.47813/rosnio-ii.2023.8.263-266.

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The article demonstrates the prospects for using a new experimental approach for the detection of the fungus Cercospora Sojina Hara. The main principle underlying the presented method is a combination of two technologies: RPA (Recombinase polymerase amplification) and CRISPR/Cas12a. RPA - is an alternative to classical PCR, with features in the form of a faster reaction rate and its passage under isothermal conditions. Using RPA technology will reduce amplification to 15-30 minutes. Amplified genomic DNA can be detected fluorescently labeled with CRISPR/Cas12a. The difficulties of this method lie in the selection of specific primers and the selection of spacers for the guide RNA within the amplicon. As a result of the work carried out, using the primer3 plugin on the Unipro Ugene platform, it was possible to select a specific pair of primers that would make it possible to identify this particular phytopathogen. The genome spacer was identified in the ChopChop web toolkit. The resulting primer pair and spacer having complementarity exclusively to the CP036215 locus in Cercospora Sojina H.

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Pernak, Elizaveta, Mariia Vladimirova, Viktoria Muntyan, Alexey Afonin und Marina Roumiantseva. „ANALYSIS OF CRISPR CASSETTE ELEMENTS IN NATIVE ISOLATES OF SINORHIZOBIUM MELILOTI ISOLATED IN THE CENTRAL ASIAN ORIGIN OF PLANT DIVERSITY“. In 23rd SGEM International Multidisciplinary Scientific GeoConference 2023. STEF92 Technology, 2023. http://dx.doi.org/10.5593/sgem2023/6.1/s25.13.

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CRISPR-Cas adaptive immunity systems of prokaryotes have been identified in 42% of bacteria and 85% of archaea [1] and are actively used in genetic engineering. However, in nitrogen-fixing nodule bacteria required for agriculture, these systems are very poorly characterized. Therefore, in this study, we evaluated the diversity of CRISPR-Cas systems in the genomes of 39 isolates of alfalfa nodule bacteria Sinorhizobium meliloti recovered from soil samples from Kazakhstan and Turkmenistan, which belong to the Central Asian origion of plants diversity. The diversity of detected CRISPR elements was assessed by changes in the size of the PCR amplified DNA fragments. As a result, the presence of 7 species-specific CRISPR loci (CR1- CR7) was shown for each of 39 isolates, of which three (CR4, CR5 and CR6) were selected for further analysis. It was shown that structural changes in the analyzed CRISPR loci sequences were significantly 2-fold more frequent in strains from Turkmenistan than in strains from Kazakhstan (?2=4.07, P less than 0.05). The most frequent changes in the size of the amplified sequences were detected at the CR6 locus (frequency 0.39), and 7.5-fold less frequent at the CR4 locus. Analysis of the nucleotide sequences of the loci for which changes in the size of amplified fragments were detected showed that at the CR4 locus one spacer was replaced with another; at the CR6 locus a deletion of the spacer and one direct repeat were involved. No changes were detected in the sequences of the CR-5 locus. The results suggest that strains in which changes in cassette compositions have occurred may have significant differences in resistance to lytic phages. The possibility of directionally modulating the spacers sequence in the CRISPR loci of nodule bacteria used in bioproducts may undoubtedly be of interest for agrobiotechnology.

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Muhlbauer, Rachel L., Kristen Turner, Finn Hiatt, Robert Dyer, Jeff Slostad und Robert P. Hoyt. „CRISSP: Customizable, Recyclable Launch Packaging“. In AIAA SPACE 2016. Reston, Virginia: American Institute of Aeronautics and Astronautics, 2016. http://dx.doi.org/10.2514/6.2016-5243.

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Tsafoulia, Loukia, und Severino Alfonso. „Transient Spaces: Building Community in Crisis Contexts“. In 2018 ACSA International Conference. ACSA Press, 2018. http://dx.doi.org/10.35483/acsa.intl.2018.1.

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Transiency no longer appears as a condition of exception, but rather as the predominant mode of existence in many parts of the world. The increased tension across and beyond national borders and territorial divisions has drawn the attention of designers across the globe and densified our reflections on questions of identity, equality, politics and economic exchange, expanding the reach of design from the realm of physical forms, into modes of interaction in social spaces. Planners and architects are being challenged to create infrastructural systems and new spatial structures of unparalleled resilience and elasticity. The paper presents part our research on the refugee crisis in the context of Greece, intertwined with the process and the experience gained as part of an advanced design studio Loukia Tsafoulia developed and taught during spring 2017 at the City College of New York. After its conclusion, the studio triggered an international call for contributions and it is currently under development for a book publication titled Transient Spaces, that explores conditions of impermanence and aims to stimulate conversations on issues of belonging and displacement.

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Baryshev, Yurij. „Physics of Gravitational Interaction: Geometry of Space or Quantum Field in Space“. In 1st CRISIS IN COSMOLOGY CONFERENCE, CCC-1. AIP, 2006. http://dx.doi.org/10.1063/1.2189130.

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Wu, Wei-Zhi, und You-Hong Xu. „Rough approximations of intuitionistic fuzzy sets in crisp approximation spaces“. In 2010 Seventh International Conference on Fuzzy Systems and Knowledge Discovery (FSKD). IEEE, 2010. http://dx.doi.org/10.1109/fskd.2010.5569662.

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Stranix, Katie. „Building Resiliency Through Small Scale Restorative Spaces“. In 2022 AIA/ACSA Intersections Research Conference. ACSA Press, 2022. http://dx.doi.org/10.35483/acsa.aia.inter.22.12.

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In the year prior to the pandemic, 1 in 3 adolescents in the US reported experiencing persistent feelings of isolation and anxiety, a 40% increase since 20091. Schools around the country recognize their role in assisting with this crisis and have increased their mental health services and staff. Although beneficial, these services require spatial compliments to increase their effectiveness and reach. Large scale renovations or entirely new constructions are often needed but require years of planning and substantial budgets to implement. How can we immediately begin to spatially address this crisis? Small scale interventions can be quickly designed and implemented with minimal disruptions to school systems, structures, and budgets. Acting together as a supportive network, these spaces can provide students, staff and faculty with accessible, restorative spaces that prioritize and support mental health. By providing schools and other shared spaces of our built environment with this type of micro restorative space, we hope to build resiliency through small scale, accessible and restorative interventions with the goal of maintaining and improving individual and community health and wellbeing. This paper focuses on the design, construction, and effectiveness of a small-scale restorative space within a main academic building on the grounds of a public university.

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Tsafoulia, Loukia. „Transient Spaces: Building Community in Crisis Contexts Project“. In 2018 ACSA International Conference. ACSA Press, 2018. http://dx.doi.org/10.35483/acsa.intl.2018.2.

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Transiency no longer appears as a condition of exception, but rather as the predominant mode of existence. The increased tension across and beyond national borders and territorial divisions has drawn the attention of designers across the globe and densified our reflections on questions of identity, equality, politics and economic exchange, expanding the reach of design from the realm of physical forms, into modes of interaction in social spaces. The project presents design outcomes from the research conducted and methods employed during the advanced architectural design studio I developed and taught at the CCNY during 2017. The studio triggered an international call for contributions and it is currently under development for a book publication that explores the concept of impermanence in global contemporary society and aims to stimulate conversations about the potential of a new public realm.

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Bachtarzi, Rym Mouni, Djamel Alkama und Hana Salah-Salah. „Urban Public Space in The Context of a New Era, Case of Annaba City-Algeria“. In 4th International Conference of Contemporary Affairs in Architecture and Urbanism – Full book proceedings of ICCAUA2020, 20-21 May 2021. Alanya Hamdullah Emin Paşa University, 2021. http://dx.doi.org/10.38027/iccaua2021199n3.

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Urban public space is an essential element of the urban environment that contributes to the quality of urban life called to create a certain dynamic, places of life, places of meeting, of use, of conviviality, and of social cohesion, providing multiple environmental and health benefits. This urban space can become even more critical in times of crisis such as the current COVID-19 pandemic, where citizens face increased health and economic stress. As part of our study, we conducted questionnaire surveys during the period in which restrictive measures were imposed in response to the pandemic in Annaba city- Algeria, which has many public urban spaces (gardens, green spaces, squares…), these surveys were intended for users on their attendance and their perception of these spaces since the onset of this pandemic COVID-19, and the challenges they see for the future. Therefore, the findings of our study demonstrated the footprint of COVID-19 on urban spaces and their frequentation, as well as preventive measures in order to gain insight into urban planning and conception.

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Lasmane, Skaidrīte. „Including the Emotional Potential of Literature in Post-crisis Education“. In 80th International Scientific Conference of the University of Latvia. University of Latvia Press, 2022. http://dx.doi.org/10.22364/htqe.2022.73.

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Contemporary situational circumstances, with the global Covid-19 pandemic crisis and the ongoing war that has resulted from Russia’s invasion of Ukraine, have brought about social, cultural, and psychological transformations that are, as of yet, little understood but already affecting different aspects of the contemporary school learning processes. Rational, analytical, cognitive, reflexive, and emotional experience are needed to ensure that difficulties within the crisis ecosystem do not cause a lessening of the human emotional experience in difficult times. Diverse emotional experiences are especially needed, the supply of which is reduced by both the limitations of interactivity imposed by the specifics of the media information space, which mostly reflects the realities of the crisis and are predominantly negative. In the face of this protracted crisis and the implications of new communication technologies, the article explores some ways to manage emotional experiences, so as not to lose sight of the diversity of human relations. It looks to address how we can compensate for the minimization of diverse emotional experience in teaching and learning in situations of social crises. The article pays attention to the potential role of literature as a way to build sustainable post-crisis social relationships. It proposes to reevaluate the role of literature in education and explore its use not only as a cognitive source for rational and critical thinking but its potential for cultivating moral emotions that enhance social solidarity and civility. The case studies it presents evaluate the interpretation and misinterpretation of some classical works of Latvian literature in schools and beyond, in the media and society.

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Berichte der Organisationen zum Thema "CRISPR spacers"

Castilleja Vargas, Liliana, Daniel Hernaiz, Alejandro Rasteletti und Eduardo Borensztein. International Crises and Policy Responses in the Southern Cone. Inter-American Development Bank, Oktober 2012. http://dx.doi.org/10.18235/0011434.

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This document analyzes the patterns of fiscal and monetary policy in five economies of the Latin American Southern Cone (Argentina, Brazil, Chile, Paraguay and Uruguay) during four episodes of international crises: 1994, 1997-1999, 2001 and 2008. In contrast with earlier episodes when most countries in the region applied procyclical fiscal and monetary policies, the response of the five countries to the 2008 crisis was countercyclical. On the fiscal side, countries had a larger fiscal space in 2008 to implement a countercyclical policy thanks to increases in primary balances, improvements in sustainability indicators and a reduction in financing needs in the years before the crisis. On the monetary side, the capacity to implement countercyclical policy was supported by a shift towards more flexible exchange rate regimes, and monetary policy regimes that enhanced transparency, reduced price volatility and increased the credibility of monetary authorities. For countries to be able to implement countercyclical responses in future crisis episodes, the fiscal authorities should reverse expansionary measures during good times and Central Banks should meet their inflation targets to preserve fiscal space and anchor inflation expectations.

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Molloy, James P. Air and Space Expeditionary Force Crisis Action Leadership for Commanders. Fort Belvoir, VA: Defense Technical Information Center, April 2004. http://dx.doi.org/10.21236/ada434089.

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Cuerden, Richard, Mary Williams, Jeanne Breen, Dan Campsal, Suzy Charman, David G. Davies, Nick Reed und Sarah Simpson. Safe Roads for All. TRL, August 2021. http://dx.doi.org/10.58446/ohss3066.

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It calls on UK Government to publish, with urgency, a Safe and Healthy Mobility Strategy and Action Plan for roads and civic spaces across the UK that is based on Safe System solutions; and for Government to place this strategy and action plan at the heart of its transport policy decisions to save people and the planet. This report proposes goals, work areas, and priority actions for the strategy and action plan. Safe and healthy mobility means we get around on roads and around our civic spaces (the spaces between our buildings) in ways that: prevent death and serious injury from road crashes; prevent death and illness from air pollution and inactivity; and achieve decarbonisation to tackle the climate crisis. We enable people to move around in active ways (walking, cycling) and we enable the safe, clean, and green use of vehicles too; to move our goods, deliver services, or move people, including by public transport.

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Rebucci, Alessandro, Andrew Powell und Eduardo Fernández-Arias. The Multilateral Response to the Global Crisis: Rationale, Modalities, and Feasibility. Inter-American Development Bank, Juni 2009. http://dx.doi.org/10.18235/0010916.

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The paper reviews the case for a strong multilateral response to the global crisis in emerging markets (EMs). It discusses modalities and feasibility of intervention and its associated risks, depending on country circumstances of fiscal space and liquidity needs. The specific role of Multilateral Development Banks (MDBs) in ensuring the development effectiveness of the fiscal response is also discussed. The paper concludes by highlighting the international financial architecture issues raised by the global crisis that cannot be addressed immediately but will need to be dealt with once the current crisis has been tamed.

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Romer, Christina, und David Romer. Fiscal Space and the Aftermath of Financial Crises: How It Matters and Why. Cambridge, MA: National Bureau of Economic Research, April 2019. http://dx.doi.org/10.3386/w25768.

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Abed, Dana, Rihab Sawaya und Nadim Tabbal. Analyzing Voter Turnout in Lebanon: Political Change in Times of Crisis. Oxfam, April 2022. http://dx.doi.org/10.21201/2022.8823.

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In May 2022, Lebanon is hosting its first parliamentary elections since the popular uprising of October 2019, when massive protests took place to denounce the current ruling elites. This research looks at voter turnout and behavior on the eve of the elections and examines the will for political change. It argues that in the current Lebanese context, there needs to be further political awareness-raising, and campaigns should be more inclusive of women and the queer community. Independent campaigns should focus on developing strong governing capacities that voters can trust, and create further space for civic and political engagement on the local and national levels.

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Sobhy, Hania. Reforms for Another Planet: The Global Learning Crisis, Political Drivers and Expert Views on Egypt’s Edu 2.0. Research on Improving Systems of Education (RISE), Februar 2023. http://dx.doi.org/10.35489/bsg-rise-2023/pe06.

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This study examines four key reform policies and programs of the Indian education system: DPEP (1994), SSA (2000), RTE (2010) and NEP (2020). Each of these has a rich and nuanced body of research to which our study contributes a unique tracing of the key ideas, debates, and stakeholders. This longue durée of reform will shed light on the histories of current reform options and if they will indeed meet the call of the times to address the learning crisis. We use the political settlements approach to understand the contestation of ideas and actors which finally determine the policy design and the implementation strategies. We unpack the iterative dynamics between ideas and actors to highlight why our reform design looks the way it does and what kind of spaces exist for transformative change particularly to ensure learning.

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Trapani, Paola. Collaborative Housing as a Response to the Housing Crisis in Auckland. Unitec ePress, Juli 2018. http://dx.doi.org/10.34074/ocds.0821.

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According to future projections based on current demographic growth trends, Auckland’s population will reach two million in 2033. Since the city is already afflicted by a serious housing crisis, at the beginning of 2017 the newly elected Mayor Phil Goff set up a task force. Formed by representatives of various stakeholders, it was given the task of producing a report with strategic and tactical guidelines to mitigate the situation. Unitec researchers were invited to respond to the report, which came out at the end of 2017, in the form of three think pieces towards the Building Better Homes, Towns and Cities National Science Challenge. This paper is a new iteration of one of these think pieces, focused on collaborative living, and expands on the new role that designers should play in this field. Its ideological position is that the house cannot and should not be considered as a commodity on the free market; nor should focus solely be on bringing down prices by increasing the number of houses on offer. Over time, housing might evolve to being more about social (use) value than exchange value. Other models of the production and consumption of household goods are documented throughout the world as alternatives to mainstream market logic, using collective procurement mechanisms to cut construction and marketing costs with savings of up to 30%. These experiments, not limited to achieving financially sustainable outcomes, are linked to new social practices of collaboration between neighbours. The sharing of spaces and equipment to complement private housing units also leads to social and environmental sustainability.

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Montiel, Peter J., und Eduardo Fernández-Arias. Crisis Response in Latin America: Is the "Rainy Day" at Hand? Inter-American Development Bank, Juni 2009. http://dx.doi.org/10.18235/0010915.

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This paper examines the countercyclical policy options available to Latin American countries in the face of the current global economic crisis, concluding that most of the major countries in the region appear to possess the fiscal space (as measured by credible fiscal sustainability and debt headroom) to run prudent countercyclical fiscal deficits. Those countries should undertake a constrained fiscal expansion focused on productive public spending and financed by rainy day fundslarge stocks of foreign exchange reserves that they have accumulated during recent yearsrather than by market borrowing. The recent surge in multilateral financial activity to alleviate market illiquidity, whether intended for reserve or budget support, strengthens the case for this policy prescription: with multilateral support, the appropriate policy response is more expansionary, and its financing is less reliant on market borrowing.

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Bonomo, Marco, Claudio R. Frischtak und Paulo Ribeiro. Public Investment and Fiscal Crisis in Brazil: Finding Culprits and Solutions. Inter-American Development Bank, April 2021. http://dx.doi.org/10.18235/0003199.

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We investigate the relation between existing fiscal rules and investments in the context of a fiscal crisis in Brazil. We analyze existing fiscal rules at national and subnational levels, their enforcement, and proposed alternatives. Using narrative analysis, case studies, interviews, empirical estimation, and model simulations, we conclude that public investment is not closely related to fiscal rules in Brazil but is mainly determined by fiscal conditions both at national and subnational (state) levels. It is the steady increase of personnel expenditures in real terms that underlies the fiscal deterioration of the last decade, despite the existence of fiscal rules devised to prevent it. We argue that a constitutional rule limiting subnationals personnel expenditures to 50 percent of net revenues, triggering adjustment measures when reaching 47.5 percent, would be an effective instrument for subnational fiscal management, opening fiscal space for increasing investments. At the national level, despite the existence of several fiscal rules, the only effective fiscal anchor is the primary expenditure ceiling introduced in 2016, which has successfully curbed expenditures, including those of the judiciary and legislature.

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