Journal articles on the topic 'Gene Editing (CRISPR/Cas9)'
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1
Paul Strickland, Skylar. "CRISPR-Cas9: Gene Editing." International Journal of Science and Research (IJSR) 12, no. 6 (June 5, 2023): 2439–42. http://dx.doi.org/10.21275/sr23624231215.
2
Yang, Jiayi. "Applications of the CRISPR-Cas9 system in cancer models." Theoretical and Natural Science 21, no. 1 (December 20, 2023): 28–33. http://dx.doi.org/10.54254/2753-8818/21/20230804.
Abstract:
Cancer has a high mortality and prevalence rate in the world. CRISPR-Cas9 is one of the novel and most common gene-editing techniques. Compared with the first two generations of gene-editing technologies, CRISPR-Cas9 system has the advantages of easy design, low cost, high efficiency and so on. sgRNA guides Cas9 to the site of the targeted gene, and Cas9 cuts the DNA strand at that site, triggering the NHEJ or HDR mechanism so as to achieve the purpose of deletion or insertion. CRISPR-Cas9 can be combined with other factors for other purposes, such as CRISPRa, CRISPRi, and base editing. The CRISPR system now has been used extensively for research into biological mechanisms and disease treatments. Since cancer is controlled by genes, a number of researchers in recent years have looked at using the CRISPR system to treat cancer. The CRISPR technology has greatly improved our understanding of cancer and the factors that affect it, and has had a major impact on the study and treatment of cancer. CRISPR gene editing can quickly and efficiently generate gene knockouts and regulate gene expression to identify relevant genes that influence cancer growth. This review systematically introduces CRISPR-Cas9 and its application methods, delivery modes, and discusses some studies using cell lines and organoids in vitro and animal models for cancer therapy in vivo.
3
Isachenko, Nadya, Gayane Aleksanyan, Paul Diehl, and Donato Tedesco. "Abstract 2950: CRISPR/saCas9 and CRISPR/spCas9 systems for combinatorial genetic screens (CRISPR-KO, CRISPRa, CRISPRi)." Cancer Research 84, no. 6_Supplement (March 22, 2024): 2950. http://dx.doi.org/10.1158/1538-7445.am2024-2950.
Abstract:
Abstract This study explores the utilization of orthogonal CRISPR-based gene editing/modulation systems for combinatorial genetic screens using CRISPR knockout (CRISPR-KO), CRISPR activation (CRISPRa), and CRISPR interference (CRISPRi) functionalities. S. aureus (sa)Cas9 is an alternative nuclease to S. pyogenes (sp)Cas9 in scenarios where the latter cannot be used, or when multiple independent CRISPR systems need to be simultaneously expressed in the same cell. In this study we set out to explore the feasibility of utilizing different combinations of saCas9 and spCas9 CRISPR systems to achieve the simultaneous inactivation (via CRISPR-KO or CRISPRi) and transactivation (via CRISPRa) of different target genes in the same host cell. For this purpose, a complete set of tools for CRISPR/saCas9 gene editing and gene modulation was developed, compatible with CRISPR/spCas9 co-expression. Specifically, we developed and validated optimized saCas9 and sg(sa)RNA lentiviral and AAV vectors, dual expression (sp)/(sa)sgRNA lentiviral library vectors, as well as (sa)CRISPR-KO, (sa)CRISPRi, (sa)CRISPRa fluorescence-based activity kits for the functional validation of saCas9 expressing cell lines. Results demonstrating the feasibility of the orthogonal screens will be presented, with different combinations of CRISPR-KO, CRISPRa, and CRISPRi systems in multiple cell lines. Citation Format: Nadya Isachenko, Gayane Aleksanyan, Paul Diehl, Donato Tedesco. CRISPR/saCas9 and CRISPR/spCas9 systems for combinatorial genetic screens (CRISPR-KO, CRISPRa, CRISPRi) [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 2950.
4
Gong, Chongzhi, Shengchan Huang, Rentao Song, and Weiwei Qi. "Comparative Study between the CRISPR/Cpf1 (Cas12a) and CRISPR/Cas9 Systems for Multiplex Gene Editing in Maize." Agriculture 11, no. 5 (May 10, 2021): 429. http://dx.doi.org/10.3390/agriculture11050429.
Abstract:
Although the clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) system has been proved to be an efficient multiplex gene editing system in maize, it was still unclear how CRISPR/Cpf1 (Cas12a) system would perform for multiplex gene editing in maize. To this end, this study compared the CRISPR/Cpf1 system and CRISPR/Cas9 system for multiplex gene editing in maize. The bZIP transcription factor Opaque2 (O2) was used as the target gene in both systems. We found that in the T0 and T1 generations, the CRISPR/Cpf1 system showed lower editing efficiency than the CRISPR/Cas9 system. However, in the T2 generation, the CRISPR/Cpf1 system generated more types of new mutations. While the CRISPR/Cas9 system tended to edit within the on-target range, the CRISPR/Cpf1 system preferred to edit in between the targets. We also found that in the CRISPR/Cpf1 system, the editing efficiency positively correlated with the expression level of Cpf1. In conclusion, the CRISPR/Cpf1 system offers alternative choices for target-site selection for multiplex gene editing and has acceptable editing efficiency in maize and is a valuable alternative choice for gene editing in crops.
5
Dowdy, Steven F. "Controlling CRISPR-Cas9 Gene Editing." New England Journal of Medicine 381, no. 3 (July 18, 2019): 289–90. http://dx.doi.org/10.1056/nejmcibr1906886.
6
Wu, Yirui. "The Development of Gene Editing Technology and Controversial Issues: A Discussion." Highlights in Science, Engineering and Technology 91 (April 15, 2024): 123–30. http://dx.doi.org/10.54097/6gj0tk11.
Abstract:
The fields of genetics and biomedicine stand to benefit greatly from the innovative technique known as gene editing technology, especially with the potential uses of CRISPR-Cas9.the creation of crops resistant to disease, the treatment of genetic illnesses, and the advancement of biomedical research in fields like sickle-cell anemia and rice gene editing are all made possible by CRISPR-Cas9's capacity to precisely pinpoint and edit specific genes. Despite being beneficial to humanity, the quick development of human gene editing technology has also brought up ethical, legal, societal, and technical issues. Its management, informed consent, privacy, and social equality are among the issues that require considerable attention.In an effort to give the scientific community something to think about, this article examines the state of gene editing today, its potential for the future, and the debates surrounding its application. It concentrates on the way the CRISPR/Cas system works, how CRISPER/Cas9 gene editing is used, and the range of problems it has brought up.
7
Yang, Lan, Hao Li, Yao Han, Yingjie Song, Mingchen Wei, Mengya Fang, and Yansong Sun. "CRISPR/Cas9 Gene Editing System Can Alter Gene Expression and Induce DNA Damage Accumulation." Genes 14, no. 4 (March 27, 2023): 806. http://dx.doi.org/10.3390/genes14040806.
Abstract:
Clustered regularly interspaced short palindromic repeats (CRISPR) and the associated protein (Cas) gene editing can induce P53 activation, large genome fragment deletions, and chromosomal structural variations. Here, gene expression was detected in host cells using transcriptome sequencing following CRISPR/Cas9 gene editing. We found that the gene editing reshaped the gene expression, and the number of differentially expressed genes was correlated with the gene editing efficiency. Moreover, we found that alternative splicing occurred at random sites and that targeting a single site for gene editing may not result in the formation of fusion genes. Further, gene ontology and KEGG enrichment analysis showed that gene editing altered the fundamental biological processes and pathways associated with diseases. Finally, we found that cell growth was not affected; however, the DNA damage response protein—γH2AX—was activated. This study revealed that CRISPR/Cas9 gene editing may induce cancer-related changes and provided basic data for research on the safety risks associated with the use of the CRISPR/Cas9 system.
8
Zhou, Junming, Xinchao Luan, Yixuan Liu, Lixue Wang, Jiaxin Wang, Songnan Yang, Shuying Liu, Jun Zhang, Huijing Liu, and Dan Yao. "Strategies and Methods for Improving the Efficiency of CRISPR/Cas9 Gene Editing in Plant Molecular Breeding." Plants 12, no. 7 (March 28, 2023): 1478. http://dx.doi.org/10.3390/plants12071478.
Abstract:
Following recent developments and refinement, CRISPR-Cas9 gene-editing technology has become increasingly mature and is being widely used for crop improvement. The application of CRISPR/Cas9 enables the generation of transgene-free genome-edited plants in a short period and has the advantages of simplicity, high efficiency, high specificity, and low production costs, which greatly facilitate the study of gene functions. In plant molecular breeding, the gene-editing efficiency of the CRISPR-Cas9 system has proven to be a key step in influencing the effectiveness of molecular breeding, with improvements in gene-editing efficiency recently becoming a focus of reported scientific research. This review details strategies and methods for improving the efficiency of CRISPR/Cas9 gene editing in plant molecular breeding, including Cas9 variant enzyme engineering, the effect of multiple promoter driven Cas9, and gRNA efficient optimization and expression strategies. It also briefly introduces the optimization strategies of the CRISPR/Cas12a system and the application of BE and PE precision editing. These strategies are beneficial for the further development and optimization of gene editing systems in the field of plant molecular breeding.
9
Desai, Devam, Hiral Panchal, Shivani Patel, and Ketul Nayak. "CRISPR - CAS9 GENE EDITING: A REVIEW." International Journal of Advanced Research 8, no. 10 (October 31, 2020): 1127–32. http://dx.doi.org/10.21474/ijar01/11943.
Abstract:
CRISPR is an RNA guided genome editing technique of genetic engineering which works like genetic scissors. Based on simplified version of bacterial CRISPR-Cas9 antiviral defense system. It is more accurate, faster and cost efficient than other genome editing methods. There are two components in this system: First component includes a single guide RNA (sgRNA) of system which will identify target sequence in genome and Second component will include Cas9 nuclease of system which will act as a pair of scissors to spilt the double strands of DNA. CRISPR has promising therapeutic applications. This current review focuses on mechanism, therapeutic applications, delivery systems, limitations and different approaches used for gene editing using CRISPR.
10
Preece, Roland, and Christos Georgiadis. "Emerging CRISPR/Cas9 applications for T-cell gene editing." Emerging Topics in Life Sciences 3, no. 3 (April 2, 2019): 261–75. http://dx.doi.org/10.1042/etls20180144.
Abstract:
Abstract Gene editing tools are being rapidly developed, accelerating many areas of cell and gene therapy research. Each successive gene editing technology promises increased efficacy, improved specificity, reduced manufacturing cost and design complexity; all of which are currently epitomised by the clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated protein (Cas9) platform. Since its conceptualisation, CRISPR-based gene editing has been applied to existing methodologies and has further allowed the exploration of novel avenues of research. Implementation of CRISPR/Cas9 has been instrumental to recent progress in the treatment of cancer, primary immunodeficiency, and infectious diseases. To this end, T-cell therapies have attempted to harness and redirect antigen recognition function, and through gene editing, broaden T-cell targeting capabilities and enhance their potency. The purpose of this review is to provide insights into emerging applications of CRISPR/Cas9 in T-cell therapies, to briefly address concerns surrounding CRISPR-mediated indel formation, and to introduce CRISPR/Cas9 base editing technologies that hold vast potential for future research and clinical translation.
11
Yang, Caiting, Yu Lei, Tinglin Ren, and Mingze Yao. "The Current Situation and Development Prospect of Whole-Genome Screening." International Journal of Molecular Sciences 25, no. 1 (January 4, 2024): 658. http://dx.doi.org/10.3390/ijms25010658.
Abstract:
High-throughput genetic screening is useful for discovering critical genes or gene sequences that trigger specific cell functions and/or phenotypes. Loss-of-function genetic screening is mainly achieved through RNA interference (RNAi), CRISPR knock-out (CRISPRko), and CRISPR interference (CRISPRi) technologies. Gain-of-function genetic screening mainly depends on the overexpression of a cDNA library and CRISPR activation (CRISPRa). Base editing can perform both gain- and loss-of-function genetic screening. This review discusses genetic screening techniques based on Cas9 nuclease, including Cas9-mediated genome knock-out and dCas9-based gene activation and interference. We compare these methods with previous genetic screening techniques based on RNAi and cDNA library overexpression and propose future prospects and applications for CRISPR screening.
12
Li, Yunhe, and Boyang Zhang. "Cancer Research Using Crispr/Cas9 Gene Editing Technology." Highlights in Science, Engineering and Technology 45 (April 18, 2023): 291–95. http://dx.doi.org/10.54097/hset.v45i.7443.
Abstract:
Gene editing technology has completely transformed the life sciences and medical treatment, because it makes precise and stable genetic alterations to genomic sequences possible. Among gene editing technologies, CRISPR system is now the most employed one. CRISPR technology is currently often used in the medical field including cancer study as well as genome editing in a number of animals. Besides, the CRISPR system has been widely used in agriculture, industry and other fields, such as crop breeding, industrial microbial design, viral nucleic acid detection, etc. In the framework of people’s knowledge of cancer research and genome editing technology, this paper focuses on the adhibition of CRISPR/Cas9 in cancer modeling, high throughput genetic analysis of tumor cell metastasis-related genes, as well as cancer treatment. It indicates that CRISPR/Cas9 techniques can be a promising tool in cancer diagnostics and therapeutics. Although CRISPR / Cas9 technology has limitations such as being off-target, this technology undoubtedly has great potential in cancer or other diseases in the future.
13
Liu, Bin, Siwei Chen, Anouk La Rose, Deng Chen, Fangyuan Cao, Martijn Zwinderman, Dominik Kiemel, Manon Aïssi, Frank J. Dekker, and Hidde J. Haisma. "Inhibition of histone deacetylase 1 (HDAC1) and HDAC2 enhances CRISPR/Cas9 genome editing." Nucleic Acids Research 48, no. 2 (December 4, 2019): 517–32. http://dx.doi.org/10.1093/nar/gkz1136.
Abstract:
Abstract Despite the rapid development of CRISPR/Cas9-mediated gene editing technology, the gene editing potential of CRISPR/Cas9 is hampered by low efficiency, especially for clinical applications. One of the major challenges is that chromatin compaction inevitably limits the Cas9 protein access to the target DNA. However, chromatin compaction is precisely regulated by histone acetylation and deacetylation. To overcome these challenges, we have comprehensively assessed the impacts of histone modifiers such as HDAC (1–9) inhibitors and HAT (p300/CBP, Tip60 and MOZ) inhibitors, on CRISPR/Cas9 mediated gene editing efficiency. Our findings demonstrate that attenuation of HDAC1, HDAC2 activity, but not other HDACs, enhances CRISPR/Cas9-mediated gene knockout frequencies by NHEJ as well as gene knock-in by HDR. Conversely, inhibition of HDAC3 decreases gene editing frequencies. Furthermore, our study showed that attenuation of HDAC1, HDAC2 activity leads to an open chromatin state, facilitates Cas9 access and binding to the targeted DNA and increases the gene editing frequencies. This approach can be applied to other nucleases, such as ZFN and TALEN.
14
Haas, Amanda. "DNA-Free CRISPR-Cas9 Gene Editing." Genetic Engineering & Biotechnology News 36, no. 17 (October 2016): 16–17. http://dx.doi.org/10.1089/gen.36.17.07.
15
Rabaan, Ali A., Hajir AlSaihati, Rehab Bukhamsin, Muhammed A. Bakhrebah, Majed S. Nassar, Abdulmonem A. Alsaleh, Yousef N. Alhashem, et al. "Application of CRISPR/Cas9 Technology in Cancer Treatment: A Future Direction." Current Oncology 30, no. 2 (February 6, 2023): 1954–76. http://dx.doi.org/10.3390/curroncol30020152.
Abstract:
Gene editing, especially with clustered regularly interspaced short palindromic repeats associated protein 9 (CRISPR-Cas9), has advanced gene function science. Gene editing’s rapid advancement has increased its medical/clinical value. Due to its great specificity and efficiency, CRISPR/Cas9 can accurately and swiftly screen the whole genome. This simplifies disease-specific gene therapy. To study tumor origins, development, and metastasis, CRISPR/Cas9 can change genomes. In recent years, tumor treatment research has increasingly employed this method. CRISPR/Cas9 can treat cancer by removing genes or correcting mutations. Numerous preliminary tumor treatment studies have been conducted in relevant fields. CRISPR/Cas9 may treat gene-level tumors. CRISPR/Cas9-based personalized and targeted medicines may shape tumor treatment. This review examines CRISPR/Cas9 for tumor therapy research, which will be helpful in providing references for future studies on the pathogenesis of malignancy and its treatment.
16
Prodipto Bishnu, Angon. "Role of CRISPR-Cas9 in agricultural science." Archives of Food and Nutritional Science 6, no. 1 (December 23, 2022): 090–91. http://dx.doi.org/10.29328/journal.afns.1001043.
Abstract:
Clustered regularly interspaced short palindromic repeat (CRISPR), a potent gene-editing tool was found in 2012. CRISPR is a genetic engineering technique that enables genome editing in living creatures and is based on the bacterial CRISPR-Cas9 antiviral defense mechanism. It is simpler, less expensive, and more accurate than previous gene editing techniques. It also has a wide range of valuable uses, including improving crops and treating genetic diseases. Plant science has benefited more from the CRISPR/Cas9 editing technique than medical science. CRISPR/Cas9 has been used in a range of crop-related research and development domains, including disease resistance, plant development, abiotic tolerance, morphological development, secondary metabolism, and fiber creation, as a well-developed cutting-edge biotechnology technique. This paper summarized the role of the CRISPR-CAS9 tool in modern agricultural science.
17
Kryštofová, Svetlana. "CRISPR/Cas in genome defense and gene editing." Acta Chimica Slovaca 9, no. 1 (April 1, 2016): 68–74. http://dx.doi.org/10.1515/acs-2016-0012.
Abstract:
AbstractTargeted genome editing using engineered nucleases such as ZFNs and TALENs has been rapidly replaced by the CRISPR/Cas9 (clustered, regulatory interspaced, short palindromic/ CRISPR-associated nuclease) system. CRISPR/Cas9 technology represents a significant improvement enabling a new level of targeting, efficiency and simplicity. Gene editing mediated by CRISPR/Cas9 has been recently used not only in bacteria but in many eukaryotic cells and organisms, from yeasts to mammals. Other modifications of the CRISPR-Cas9 system have been used to introduce heterologous domains to regulate gene expressions or label specific loci in various cell types. The review focuses not only on native CRISPR/Cas systems which evolved in prokaryotes as an endogenous adaptive defense mechanism against foreign DNA attacks, but also on the CRISPR/Cas9 adoption as a powerful tool for site-specific gene modifications in fungi, plants and mammals.
18
Doench, John G. "CRISPR/Cas9 gene editing special issue." FEBS Journal 283, no. 17 (September 2016): 3160–61. http://dx.doi.org/10.1111/febs.13823.
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America Fernanda Acosta-Soto, Diana Marisol López-Díaz, Jehosafat Esquivel-Ramírez, Joana Mora-Soriano, and Brissia Lazalde- Medina. "Fundamentals of CRISPR-Cas9: Gene-editing technology and basic." GSC Advanced Research and Reviews 20, no. 1 (July 30, 2024): 042–49. http://dx.doi.org/10.30574/gscarr.2024.20.1.0223.
Abstract:
The CRISPR/Cas9 system provides a robust and multiplexable genome editing tool, enabling researchers to precisely manipulate specific genomic elements and facilitating the elucidation of target gene function in biology and diseases. CRISPR/Cas9 consists of a nonspecific Cas9 nuclease and a set of programmable sequence-specific CRISPR RNA (crRNA), which can guide Cas9 to cleave DNA and generate double-strand breaks at target sites. Subsequent cellular DNA repair processes lead to desired insertions, deletions, or substitutions at target sites. The particularity of CRISPR/Cas9-mediated DNA cleavage requires target sequences matching crRNA and a protospacer adjacent motif located downstream of target sequences. Here, we review the molecular mechanism, applications, and challenges of CRISPR/Cas9-mediated genome editing and the clinical therapeutic potential of CRISPR/Cas9 in the future.
20
Liu, Hua, Wendan Chen, Yushu Li, Lei Sun, Yuhong Chai, Haixia Chen, Haochen Nie, and Conglin Huang. "CRISPR/Cas9 Technology and Its Utility for Crop Improvement." International Journal of Molecular Sciences 23, no. 18 (September 9, 2022): 10442. http://dx.doi.org/10.3390/ijms231810442.
Abstract:
The rapid growth of the global population has resulted in a considerable increase in the demand for food crops. However, traditional crop breeding methods will not be able to satisfy the worldwide demand for food in the future. New gene-editing technologies, the most widely used of which is CRISPR/Cas9, may enable the rapid improvement of crop traits. Specifically, CRISPR/Cas9 genome-editing technology involves the use of a guide RNA and a Cas9 protein that can cleave the genome at specific loci. Due to its simplicity and efficiency, the CRISPR/Cas9 system has rapidly become the most widely used tool for editing animal and plant genomes. It is ideal for modifying the traits of many plants, including food crops, and for creating new germplasm materials. In this review, the development of the CRISPR/Cas9 system, the underlying mechanism, and examples of its use for editing genes in important crops are discussed. Furthermore, certain limitations of the CRISPR/Cas9 system and potential solutions are described. This article will provide researchers with important information regarding the use of CRISPR/Cas9 gene-editing technology for crop improvement, plant breeding, and gene functional analyses.
21
Cai, Ruijie, Runyu Lv, Xin’e Shi, Gongshe Yang, and Jianjun Jin. "CRISPR/dCas9 Tools: Epigenetic Mechanism and Application in Gene Transcriptional Regulation." International Journal of Molecular Sciences 24, no. 19 (October 3, 2023): 14865. http://dx.doi.org/10.3390/ijms241914865.
Abstract:
CRISPR/Cas9-mediated cleavage of DNA, which depends on the endonuclease activity of Cas9, has been widely used for gene editing due to its excellent programmability and specificity. However, the changes to the DNA sequence that are mediated by CRISPR/Cas9 affect the structures and stability of the genome, which may affect the accuracy of results. Mutations in the RuvC and HNH regions of the Cas9 protein lead to the inactivation of Cas9 into dCas9 with no endonuclease activity. Despite the loss of endonuclease activity, dCas9 can still bind the DNA strand using guide RNA. Recently, proteins with active/inhibitory effects have been linked to the end of the dCas9 protein to form fusion proteins with transcriptional active/inhibitory effects, named CRISPRa and CRISPRi, respectively. These CRISPR tools mediate the transcription activity of protein-coding and non-coding genes by regulating the chromosomal modification states of target gene promoters, enhancers, and other functional elements. Here, we highlight the epigenetic mechanisms and applications of the common CRISPR/dCas9 tools, by which we hope to provide a reference for future related gene regulation, gene function, high-throughput target gene screening, and disease treatment.
22
Gupta, Ajeet Kumar, Govind Mishra, Harikant Yadav, Rishabh Gupta, Abhay Singh, Jay Singh, and Piyusha Singh. "Enhancing Crop Resilience through CRISPR/Cas9-Mediated Development of Disease-Resistant Cultivars." International Journal of Environment and Climate Change 13, no. 10 (September 9, 2023): 2773–83. http://dx.doi.org/10.9734/ijecc/2023/v13i102942.
Abstract:
A major issue in agriculture is the protection of crops against diseases and pests. Plant breeding has been primarily responsible for the growth of disease-resistant cultivars. The use of gene editing techniques in plant breeding is essential for obtaining desired features. Clustered Regular Interspaced Palindromic Repeats (CRISPER)/Cas9 (CRISPR-related protein) is a new advancement in gene editing technology. It can be utilised in plant defence mechanisms against pathogen attack by recognising the bacterial immune system and destroying invasive pathogen genes. Advances in plant breeding through CRISPR/Cas9 integration have helped develop cultivars including hereditary resistance to bacterial and viral diseases. Future crop generations can acquire CRISPR/Cas9-mediated transgene resistance if the Cas9/sgRNA transgene has been isolated in the F1 generation. Cas9/sgRNA transgene separation makes CRISPR/Cas9 safe for use in plant breeding. Although CRISPR/Cas9 has proven to be a wonderful tool to revolutionize plant breeding and develop various disease resistant varieties, its effect on many plant physiological processes remains to be thoroughly investigated.
23
Xia, Kaisheng. "Application of CRISPR gene editing technology in cystic fibrosis treatment." Highlights in Science, Engineering and Technology 73 (November 29, 2023): 384–89. http://dx.doi.org/10.54097/hset.v73i.13107.
Abstract:
CRISPR-Cas9 gene editing technology is to identify the target genome sequence through artificially designed sgRNA, guide Cas9 protease, so that it can effectively cut the DNA double strand, form a double strand break, repair after damage to lead to gene knockout or knock-in, and finally achieve the purpose of modifying genomic DNA which leading CRISPR a technology can effectively repair, correct or regulate genetic mutations associated with genetic diseases, providing patients with new treatment options. However, despite the remarkable progress, CRISPR technology still faces many challenges in its clinical application, including ethics and safety. This article reviews the principle and development of CRISPR technology, as well as its application in the treatment of cystic fibrosis, and summarizes the challenges of CRISPR in clinical applications. More importantly, this article focuses on some current improvements to CRIPSR to make CRISPR gene editing technology more valuable and meaningful in the treatment of genetic diseases in the future.
24
Qiu, Yiwen, Jiaying Wang, Youhui Yao, and Ziqiu Yin. "Application prospect of CRISPR-Cas9 gene editing in epilepsy." Highlights in Science, Engineering and Technology 74 (December 29, 2023): 126–33. http://dx.doi.org/10.54097/shcr4j47.
Abstract:
Epilepsy, a chronic noncommunicable brain disorder, is characterized by abnormal electrical activity in the brain, leading to seizures and disruptions in normal brain functions. Despite various known causes, a significant proportion of epilepsy cases remain unexplained. In recent years, Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR-Cas9) technology has emerged as a powerful tool for genetic engineering. Compared with the first two generations of gene-editing technology, it has the advantages of low cost, easy design and easy operation. Utilizing CRISPR activation (CRISPRa), researchers have explored the potential of increasing the expression of genes involved in regulating synaptic interactions to control epileptic activity. Studies on transgenic mice have shown that upregulating the Kv1.1 gene (Kcnal1), which encodes for a voltage-gated potassium channel responsible for regulating neuronal excitability, can reduce seizures and improve cognitive function. Additionally, CRISPR-Cas9 has been instrumental in creating animal models to study epilepsy, providing insights into gene functions, disease mechanisms, and potential therapeutic interventions. However, further research is needed to fully explore the potential of CRISPR-based therapies for targeted treatment of epilepsy. This review systematically introduces the pathogenesis of epilepsy, including the origins and causes of epilepsy and the mechanism of seizure formation and further discusses the application of the CRISPR/Cas9 system in epilepsy.
25
Khan, Sikandar. "Recent Advancement and Innovations in CRISPR/Cas and CRISPR Related Technologies: A review." Biotechnology and Bioprocessing 2, no. 5 (June 24, 2021): 01–12. http://dx.doi.org/10.31579/2766-2314/042.
Abstract:
CRISPR genome editing technologies have been improving by every passing day. The initial CRISPR/Cas9 technologies, though emerged an improved version of genome editing in competition with TALENS and ZFNs, was nevertheless not free from technical and off-target effects. Technological improvements overtime start addressing issues with original CRISPR/Cas9 technology. The major areas of improvement targeted nucleases and delivery methods. Overtime the nuclease like Cas9 had some modifications like FokI-dCas9, Truncated guide RNAs (tru-gRNAs), Paired Cas9 nickase, Cpf1, Cas6 with Csm/Csr complex and chemically treated Cas9. In terms of delivery methods the improvements came along after almost all methods including viral methods like Recombinant Adeno Associated Viruses (rAAV), Lentivirus (LV), and bacteriophages. The review summarizes various non-viral gene delivery modes including physical methods like electroporation and chemical methods like nano particles, cell-derived membrane vesicles (CMVs) with upgraded developments. The review also compares various modes of delivering CRISPR gene editing machinery.
26
Sun, Jinyu, Jianchu Wang, Donghui Zheng, and Xiaorong Hu. "Advances in therapeutic application of CRISPR-Cas9." Briefings in Functional Genomics 19, no. 3 (November 26, 2019): 164–74. http://dx.doi.org/10.1093/bfgp/elz031.
Abstract:
Abstract Clustered regularly interspaced short palindromic repeats (CRISPR)-associated protein 9 (Cas9) is one of the most versatile and efficient gene editing technologies, which is derived from adaptive immune strategies for bacteria and archaea. With the remarkable development of programmable nuclease-based genome engineering these years, CRISPR-Cas9 system has developed quickly in recent 5 years and has been widely applied in countless areas, including genome editing, gene function investigation and gene therapy both in vitro and in vivo. In this paper, we briefly introduce the mechanisms of CRISPR-Cas9 tool in genome editing. More importantly, we review the recent therapeutic application of CRISPR-Cas9 in various diseases, including hematologic diseases, infectious diseases and malignant tumor. Finally, we discuss the current challenges and consider thoughtfully what advances are required in order to further develop the therapeutic application of CRISPR-Cas9 in the future.
27
Ding, Anqi, Zhongjin Gu, and Zihan Chen. "Application of CRISPR-Cas 9 system in cancer therapy." Highlights in Science, Engineering and Technology 74 (December 29, 2023): 302–6. http://dx.doi.org/10.54097/s50r4154.
Abstract:
There are several gene editing technologies under the background of that time, such as ZFN and TALEN which are hard to use and have high cost. On the contrary, the efficiency and accuracy of CRISPR-Cas system has attracted the attention of scientists. CRISPR is a Short palindromic repeat sequence, and it is widespread in many prokaryotes. The first CRISPR was cloned in E. coli by scientists in 1987, and several other sequences were subsequently cloned. The Cas is a nuclease, then two of them cooperate to become the CRISPR-Cas system. It is not only an acquired immune defense mechanism of prokaryotes against virus, but also a tool for gene editing. As an emerging gene editing technology in recent years, CRISPR-Cas9 technology has been widely applied to the treatment of a variety of diseases. This review focused on the application of this technology in cancer. This review systematically introduced the working principle and immune mechanism of CRISPR-Cas9 and compared CRISPR-Cas9 technology with two previous generations of gene-editing technology, current application of CRISPR-Cas9 in cancer treatment, elaborating it with relevant examples, and finally pointed out the challenges of CRISPR-Cas9 in cancer treatment, looking forward to the development of this technology in the future. The purpose of the paper is to introduce and popularize CRISPR-Cas9 technology, and to provide a direction for future cancer treatment.
28
Jinka, Chaitra. "CRISPR-Cas9 gene editing and human diseases." Bioinformation 18, no. 11 (November 30, 2022): 1081–86. http://dx.doi.org/10.6026/973206300181081.
Abstract:
CRISPR/Cas-9 mediated genome editing has recently emerged as a potential and innovative technology in therapeutic development and biomedical research. Several recent studies have been performed to understand gene modification techniques in obtaining effective ex vivo results. Generally, the disease targets for gene correction will be in specific organs, so understanding the complete potential of genomic treatment methods is essential. From such a perspective, the present review revealed the significant importance of the CRISPR/ Cas9 delivery system. Both the promising gene-editing delivery systems, such as synthetic (non-viral) and viral vector systems are discussed in this review. In addition, this paper attempted to summarize the tissue-specific and organ-specific mRNA delivery systems that provide possible research information for future researchers. Further, the major challenges of the CRISPR/Cas9 system, such as off-target delivery, immunogenicity, and limited packaging, were also elucidated. Accordingly, this review illustrated a wide range of clinical applications associated with the efficient delivery of CRISPR/ Cas9 gene-editing. Moreover, this article emphasizes the role of the CRISPR/Cas9 system in treating Intra Cerebral haemorrhage (ICH), thereby suggesting future researchers to adopt more clinical trials on this breakthrough delivery system.
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Park, Hanseul, Jaein Shin, Hwan Choi, Byounggook Cho, and Jongpil Kim. "Valproic Acid Significantly Improves CRISPR/Cas9-Mediated Gene Editing." Cells 9, no. 6 (June 10, 2020): 1447. http://dx.doi.org/10.3390/cells9061447.
Abstract:
The clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 system has emerged as a powerful technology, with the potential to generate transgenic animals. Particularly, efficient and precise genetic editing with CRISPR/Cas9 offers immense prospects in various biotechnological applications. Here, we report that the histone deacetylase inhibitor valproic acid (VPA) significantly increases the efficiency of CRISPR/Cas9-mediated gene editing in mouse embryonic stem cells and embryos. This effect may be caused through globally enhanced chromatin accessibility, as indicate by histone hyperacetylation. Taken together, our results suggest that VPA can be used to increase the efficacy of CRISPR/Cas9 in generating transgenic systems.
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Dev, Kapil, Jubeda Begum, Nasir Akbar Mir, and Rajiv Kant. "Advancements of CRISPR/Cas9 technology and its value in antiviral therapeutics." Letters In Animal Biology 1, no. 1 (September 26, 2021): 46–57. http://dx.doi.org/10.62310/liab.v1i1.60.
Abstract:
CISPR/Cas9 system is a natural immune mechanism adopted by bacteria and archaea on exposure to invading phages and plasmids. The field of genome editing has been revolutionized with the advent of CRISPR/Cas9 technology. The CRISPR/Cas9 based gene editing has offered a promising therapeutic platform for many animal and human diseases, particularly viral diseases because viruses evolve constantly and hence escape vaccine-induced immunity. The targeted genome editing by RNA-guided nucleases is rapid, easy, economical, and efficient compared to previous editing technologies. It not only helps in the direct destruction of viruses, but also helps us understand the host-virus interactions, gene functions, and develop recombinant vaccines. It has been widely experimented in the field of antiviral therapy, starting with HIV in 2013 to SARS CoV-2 recently, with a series of modifications in structure and composition of CRISPR/Cas9 and delivery mechanisms to achieve the ever-increasing promising results. Herein, this review focused on the origin of CRISPR/Cas9 system, mechanism of action, advantages over existing gene-editing tools, its progress in antiviral therapy, vaccine development, delivery approaches, and challenges faced in the application of CRISPR/Cas9.
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Singh, Sanjay. "Gene Editing Technologies: CRISPR/Cas9 and Beyond for Genetic Disease Therapy and Research." Universal Research Reports 11, no. 3 (June 30, 2024): 1–7. http://dx.doi.org/10.36676/urr.v11.i3.1280.
Abstract:
Gene editing technologies have revolutionized the field of genetic disease therapy and research. Among these, CRISPR/Cas9 stands out as a versatile tool for precisely targeting and modifying specific sequences within the genome. This paper provides an overview of CRISPR/Cas9 and other emerging gene editing technologies, discussing their potential applications in treating genetic diseases and advancing scientific research. Additionally, ethical considerations and challenges associated with gene editing are explored, along with future directions for this rapidly evolving field. Gene editing technologies have emerged as powerful tools in the field of genetic disease therapy and research, offering unprecedented precision and versatility in manipulating the genome. Among these technologies, CRISPR/Cas9 has garnered significant attention for its simplicity, efficiency, and accuracy in targeted gene modification.
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Smirnov, Kirill, Florian Weiss, Anna-Maria Hatzl, Lukas Rieder, Kjeld Olesen, Sanne Jensen, and Anton Glieder. "Comparison of CRISPR-MAD7 and CRISPR-Cas9 for Gene Disruptions in Komagataella phaffii." Journal of Fungi 10, no. 3 (March 5, 2024): 197. http://dx.doi.org/10.3390/jof10030197.
Abstract:
CRISPR (clustered regularly interspaced short palindromic repeats)-based technologies are powerful, programmable tools for site-directed genome modifications. After successful adaptation and efficient use of CRISPR-Cas9 for genome engineering in methylotrophic yeast Komagataella phaffii, a broader variety of employable endonucleases was desired to increase the experimental flexibility and to provide alternatives in case there are specific legal restrictions in industrial research due to the intellectual property rights (IPRs) of third parties. MAD7, an engineered Class 2 Type V Cas nuclease, was promoted as a royalty-free alternative for academic and industrial research and developed by Inscripta (Pleasanton, CA, USA). In this study, for the first time, CRISPR-MAD7 was used for genome editing in K. phaffii with a high gene-editing rate (up to 90%), as demonstrated for the three targeted genes coding for glycerol kinase 1 (GUT1), red fluorescence protein (DsRed), and zeocin resistance gene (Sh ble). Additionally, the genome-editing efficiencies of the CRISPR-MAD7 and CRISPR-Cas9 systems were systematically compared by targeting 259 kinase genes in K. phaffii. In this broad testing, the CRISPR-Cas9 had a higher genome-editing rate of about 65%, in comparison to the applied CRISPR-MAD7 toolbox (about 23%).
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Ding, Shuai, Jinfeng Liu, Xin Han, and Mengfan Tang. "CRISPR/Cas9-Mediated Genome Editing in Cancer Therapy." International Journal of Molecular Sciences 24, no. 22 (November 15, 2023): 16325. http://dx.doi.org/10.3390/ijms242216325.
Abstract:
The Clustered Regularly Interspaced Short Palindromic Repeats/CRISPR-associated protein 9 (CRISPR/Cas9) system, an RNA-based adaptive immune system found in bacteria and archaea, has catalyzed the development and application of a new generation of gene editing tools. Numerous studies have shown that this system can precisely target a wide range of human genes, including those associated with diseases such as cancer. In cancer research, the intricate genetic mutations in tumors have promoted extensive utilization of the CRISPR/Cas9 system due to its efficient and accurate gene editing capabilities. This includes improvements in Chimeric Antigen Receptor (CAR)-T-cell therapy, the establishment of tumor models, and gene and drug target screening. Such progress has propelled the investigation of cancer molecular mechanisms and the advancement of precision medicine. However, the therapeutic potential of genome editing remains underexplored, and lingering challenges could elevate the risk of additional genetic mutations. Here, we elucidate the fundamental principles of CRISPR/Cas9 gene editing and its practical applications in tumor research. We also briefly discuss the primary challenges faced by CRISPR technology and existing solutions, intending to enhance the efficacy of this gene editing therapy and shed light on the underlying mechanisms of tumors.
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Mu, Yulin, Chengxiao Zhang, Taihua Li, Feng-Jie Jin, Yun-Ju Sung, Hee-Mock Oh, Hyung-Gwan Lee, and Long Jin. "Development and Applications of CRISPR/Cas9-Based Genome Editing in Lactobacillus." International Journal of Molecular Sciences 23, no. 21 (October 25, 2022): 12852. http://dx.doi.org/10.3390/ijms232112852.
Abstract:
Lactobacillus, a genus of lactic acid bacteria, plays a crucial function in food production preservation, and probiotics. It is particularly important to develop new Lactobacillus strains with superior performance by gene editing. Currently, the identification of its functional genes and the mining of excellent functional genes mainly rely on the traditional gene homologous recombination technology. CRISPR/Cas9-based genome editing is a rapidly developing technology in recent years. It has been widely applied in mammalian cells, plants, yeast, and other eukaryotes, but less in prokaryotes, especially Lactobacillus. Compared with the traditional strain improvement methods, CRISPR/Cas9-based genome editing can greatly improve the accuracy of Lactobacillus target sites and achieve traceless genome modification. The strains obtained by this technology may even be more efficient than the traditional random mutation methods. This review examines the application and current issues of CRISPR/Cas9-based genome editing in Lactobacillus, as well as the development trend of CRISPR/Cas9-based genome editing in Lactobacillus. In addition, the fundamental mechanisms of CRISPR/Cas9-based genome editing are also presented and summarized.
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Huang, Wenli, Aihong Zheng, Huanhuan Huang, Zhifeng Chen, Jie Ma, Xiangxiang Li, Qiannan Liang, et al. "Effects of sgRNAs, Promoters, and Explants on the Gene Editing Efficiency of the CRISPR/Cas9 System in Chinese Kale." International Journal of Molecular Sciences 24, no. 17 (August 26, 2023): 13241. http://dx.doi.org/10.3390/ijms241713241.
Abstract:
The CRISPR/Cas9 system is extensively used for plant gene editing. This study developed an efficient CRISPR/Cas9 system for Chinese kale using multiple sgRNAs and two promoters to create various CRISPR/Cas9 vectors. These vectors targeted BoaZDS and BoaCRTISO in Chinese kale protoplasts and cotyledons. Transient transformation of Chinese kale protoplasts was assessed for editing efficiency at three BoaZDS sites. Notably, sgRNA: Z2 achieved the highest efficiency (90%). Efficiency reached 100% when two sgRNAs targeted BoaZDS with a deletion of a large fragment (576 bp) between them. However, simultaneous targeting of BoaZDS and BoaCRTISO yielded lower efficiency. Transformation of cotyledons led to Chinese kale mutants with albino phenotypes for boazds mutants and orange-mottled phenotypes for boacrtiso mutants. The mutation efficiency of 35S-CRISPR/Cas9 (92.59%) exceeded YAO-CRISPR/Cas9 (70.97%) in protoplasts, and YAO-CRISPR/Cas9 (96.49%) surpassed 35S-CRISPR/Cas9 (58%) in cotyledons. These findings introduce a strategy for enhancing CRISPR/Cas9 editing efficiency in Chinese kale.
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Meiliana, Anna, Nurrani Mustika Dewi, and Andi Wijaya. "Genome Editing with Crispr-Cas9 Systems: Basic Research and Clinical Applications." Indonesian Biomedical Journal 9, no. 1 (April 1, 2017): 1. http://dx.doi.org/10.18585/inabj.v9i1.272.
Abstract:
BACKGROUND: Recently established genome editing technologies will open new avenues for biological research and development. Human genome editing is a powerful tool which offers great scientific and therapeutic potential.CONTENT: Genome editing using the clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPRassociated protein 9 (Cas9) technology is revolutionizing the gene function studies and possibly will give rise to an entirely new degree of therapeutics for a large range of diseases. Prompt advances in the CRISPR/Cas9 technology, as well as delivery modalities for gene therapy applications, are dismissing the barriers to the clinical translation of this technology. Many studies conducted showed promising results, but as current available technologies for evaluating off-target gene modification, several elements must be addressed to validate the safety of the CRISPR/Cas9 platform for clinical application, as the ethical implication as well.SUMMARY: The CRISPR/Cas9 system is a powerful genome editing technology with the potential to create a variety of novel therapeutics for a range of diseases, many of which are currently untreatable.KEYWORDS: genome editing, CRISPR-Cas, guideRNA, DSB, ZFNs, TALEN
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Qi, Qiaoyun, Bichun Hu, Weiyu Jiang, Yixiong Wang, Jinjiao Yan, Fengwang Ma, Qingmei Guan, and Jidi Xu. "Advances in Plant Epigenome Editing Research and Its Application in Plants." International Journal of Molecular Sciences 24, no. 4 (February 8, 2023): 3442. http://dx.doi.org/10.3390/ijms24043442.
Abstract:
Plant epistatic regulation is the DNA methylation, non-coding RNA regulation, and histone modification of gene sequences without altering the genome sequence, thus regulating gene expression patterns and the growth process of plants to produce heritable changes. Epistatic regulation in plants can regulate plant responses to different environmental stresses, regulate fruit growth and development, etc. Genome editing can effectively improve plant genetic efficiency by targeting the design and efficient editing of genome-specific loci with specific nucleases, such as zinc finger nucleases (ZFNs), transcription activator-like effector nucleases (TALEN), and clustered regularly interspaced short palindromic repeats/CRISPR-associated 9 (CRISPR/Cas9). As research progresses, the CRISPR/Cas9 system has been widely used in crop breeding, gene expression, and epistatic modification due to its high editing efficiency and rapid translation of results. In this review, we summarize the recent progress of CRISPR/Cas9 in epigenome editing and look forward to the future development direction of this system in plant epigenetic modification to provide a reference for the application of CRISPR/Cas9 in genome editing.
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McAndrews, Kathleen M., Fei Xiao, Antonios Chronopoulos, Valerie S. LeBleu, Fernanda G. Kugeratski, and Raghu Kalluri. "Exosome-mediated delivery of CRISPR/Cas9 for targeting of oncogenic KrasG12D in pancreatic cancer." Life Science Alliance 4, no. 9 (July 19, 2021): e202000875. http://dx.doi.org/10.26508/lsa.202000875.
Abstract:
CRISPR/Cas9 is a promising technology for gene editing. To date, intracellular delivery vehicles for CRISPR/Cas9 are limited by issues of immunogenicity, restricted packaging capacity, and low tolerance. Here, we report an alternative, nonviral delivery system for CRISPR/Cas9 based on engineered exosomes. We show that non-autologous exosomes can encapsulate CRISPR/Cas9 plasmid DNA via commonly available transfection reagents and can be delivered to recipient cancer cells to induce targeted gene deletion. As a proof-of-principle, we demonstrate that exosomes loaded with CRISPR/Cas9 can target the mutant KrasG12D oncogenic allele in pancreatic cancer cells to suppress proliferation and inhibit tumor growth in syngeneic subcutaneous and orthotopic models of pancreatic cancer. Exosomes may thus be a promising delivery platform for CRISPR/Cas9 gene editing for targeted therapies.
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Vaishnav, Radhika A. "The emerging role of CRISPR-Cas9 in molecular oncology." International Journal of Molecular and Immuno Oncology 2, no. 2 (June 24, 2017): 45. http://dx.doi.org/10.18203/issn.2456-3994.intjmolimmunooncol20172641.
Abstract:
It is not uncommon to be curious about the recent hype surrounding the new gene editing player, Cas9, which recognizes and holds into place DNA segments known as clustered regularly interspaced short palindromic repeats (CRISPR). Together, they are known as CRISPR-Cas9 or simply “CRISPR” for brevity. The binding of Cas9 causes the CRISPR sequences to become available for editing.
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Yip, Bon Ham. "Recent Advances in CRISPR/Cas9 Delivery Strategies." Biomolecules 10, no. 6 (May 30, 2020): 839. http://dx.doi.org/10.3390/biom10060839.
Abstract:
The clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 system has revolutionized the field of gene editing. Continuous efforts in developing this technology have enabled efficient in vitro, ex vivo, and in vivo gene editing through a variety of delivery strategies. Viral vectors are commonly used in in vitro, ex vivo, and in vivo delivery systems, but they can cause insertional mutagenesis, have limited cloning capacity, and/or elicit immunologic responses. Physical delivery methods are largely restricted to in vitro and ex vivo systems, whereas chemical delivery methods require extensive optimization to improve their efficiency for in vivo gene editing. Achieving a safe and efficient in vivo delivery system for CRISPR/Cas9 remains the most challenging aspect of gene editing. Recently, extracellular vesicle-based systems were reported in various studies to deliver Cas9 in vitro and in vivo. In comparison with other methods, extracellular vesicles offer a safe, transient, and cost-effective yet efficient platform for delivery, indicating their potential for Cas9 delivery in clinical trials. In this review, we first discuss the pros and cons of different Cas9 delivery strategies. We then specifically review the development of extracellular vesicle-mediated gene editing and highlight the strengths and weaknesses of this technology.
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Zhao, Ziqi. "Principle and applications of CRISPR/Cas system." Theoretical and Natural Science 20, no. 1 (December 20, 2023): 221–26. http://dx.doi.org/10.54254/2753-8818/20/20230772.
Abstract:
With the continuous in-depth research and exploration of the CRISPR/Cas9 system by researchers, people's ability to edit biological genes is also constantly improving. In recent years, gene editing technology as a hot technology has been paid more and more attention, it has played an important role in crop breeding, disease treatment, molecular diagnosis and other aspects. Among the gene editing technologies that have been developed, the emerging CRISPR/Cas technology gene editing system has become the most widely used gene editing technology at present. Compared with the first two generations of gene-editing technologies: ZFN and TALEN, CRISPR/Cas system has the advantages of short cycle, high efficiency, low cost, and has great potential for the development of animal and plant breeding. The development of CRISPR/Cas9 technology has promoted the progress of modern life sciences. With these tools for efficient, simple, and precise genome modification, transcriptional regulation, and epigenetic editing, CRISPR systems have the potential to revolutionize genetics, medicine, and agriculture. In this review, the basic principle of CRISPR/Cas technology and its application will be introduced and illustrated.
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Hasselbeck, Sebastian, and Xinlai Cheng. "Molecular Marvels: Small Molecules Paving the Way for Enhanced Gene Therapy." Pharmaceuticals 17, no. 1 (December 27, 2023): 41. http://dx.doi.org/10.3390/ph17010041.
Abstract:
In the rapidly evolving landscape of genetic engineering, the advent of CRISPR-Cas technologies has catalyzed a paradigm shift, empowering scientists to manipulate the genetic code with unprecedented accuracy and efficiency. Despite the remarkable capabilities inherent to CRISPR-Cas systems, recent advancements have witnessed the integration of small molecules to augment their functionality, introducing new dimensions to the precision and versatility of gene editing applications. This review delves into the synergy between CRISPR-Cas technologies based specifically on Cas9 and small-molecule drugs, elucidating the pivotal role of chemicals in optimizing target specificity and editing efficiency. By examining a diverse array of applications, ranging from therapeutic interventions to agricultural advancements, we explore how the judicious use of chemicals enhances the precision of CRISPR-Cas9-mediated genetic modifications. In this review, we emphasize the significance of small-molecule drugs in fine-tuning the CRISPR-Cas9 machinery, which allows researchers to exert meticulous control over the editing process. We delve into the mechanisms through which these chemicals bolster target specificity, mitigate off-target effects, and contribute to the overall refinement of gene editing outcomes. Additionally, we discuss the potential of chemical integration in expanding the scope of CRISPR-Cas9 technologies, enabling tailored solutions for diverse genetic manipulation challenges. As CRISPR-Cas9 technologies continue to evolve, the integration of small-molecule drugs emerges as a crucial avenue for advancing the precision and applicability of gene editing techniques. This review not only synthesizes current knowledge but also highlights future prospects, paving the way for a deeper understanding of the synergistic interplay between CRISPR-Cas9 systems and chemical modulators in the pursuit of more controlled and efficient genetic modifications.
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Osadchiy, Igor S., Kamalyan O. Sophia, Karina Yu Tumashova, Pavel G. Georgiev, and Oksana G. Maksimenko. "CRISPR/Cas9 Essential Gene Editing in Drosophila." Acta Naturae 15, no. 2 (August 3, 2023): 70–74. http://dx.doi.org/10.32607/actanaturae.11874.
Abstract:
Since the addition of the CRISPR/Cas9 technology to the genetic engineering toolbox, the problems of low efficiency and off-target effects hamper its widespread use in all fields of life sciences. Furthermore, essential gene knockout usually results in failure and it is often not obvious whether the gene of interest is an essential one. Here, we report on a new strategy to improve the CRISPR/Cas9 genome editing, which is based on the idea that editing efficiency is tightly linked to how essential the gene to be modified is. The more essential the gene, the less the efficiency of the editing and the larger the number of off-targets, due to the survivorship bias. Considering this, we generated deletions of three essential genes in Drosophila: trf2, top2, and mep-1, using fly strains with previous target gene overexpression (pre-rescued genetic background).
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Seok, Heeyoung, Rui Deng, Douglas B. Cowan, and Da-Zhi Wang. "Application of CRISPR-Cas9 gene editing for congenital heart disease." Clinical and Experimental Pediatrics 64, no. 6 (June 15, 2021): 269–79. http://dx.doi.org/10.3345/cep.2020.02096.
Abstract:
Clustered regularly interspaced short palindromic repeats and CRISPR-associated protein 9 (CRISPR-Cas9) is an ancient prokaryotic defense system that precisely cuts foreign genomic DNA under the control of a small number of guide RNAs. The CRISPR-Cas9 system facilitates efficient double-stranded DNA cleavage that has been recently adopted for genome editing to create or correct inherited genetic mutations causing disease. Congenital heart disease (CHD) is generally caused by genetic mutations such as base substitutions, deletions, and insertions, which result in diverse developmental defects and remains a leading cause of birth defects. Pediatric CHD patients exhibit a spectrum of cardiac abnormalities such as septal defects, valvular defects, and abnormal chamber development. CHD onset occurs during the prenatal period and often results in early lethality during childhood. Because CRISPR-Cas9-based genome editing technology has gained considerable attention for its potential to prevent and treat diseases, we will review the CRISPR-Cas9 system as a genome editing tool and focus on its therapeutic application for CHD.
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Janik, Edyta, Marcin Niemcewicz, Michal Ceremuga, Lukasz Krzowski, Joanna Saluk-Bijak, and Michal Bijak. "Various Aspects of a Gene Editing System—CRISPR–Cas9." International Journal of Molecular Sciences 21, no. 24 (December 16, 2020): 9604. http://dx.doi.org/10.3390/ijms21249604.
Abstract:
The discovery of clustered, regularly interspaced short palindromic repeats (CRISPR) and their cooperation with CRISPR-associated (Cas) genes is one of the greatest advances of the century and has marked their application as a powerful genome engineering tool. The CRISPR–Cas system was discovered as a part of the adaptive immune system in bacteria and archaea to defend from plasmids and phages. CRISPR has been found to be an advanced alternative to zinc-finger nucleases (ZFN) and transcription activator-like effector nucleases (TALEN) for gene editing and regulation, as the CRISPR–Cas9 protein remains the same for various gene targets and just a short guide RNA sequence needs to be altered to redirect the site-specific cleavage. Due to its high efficiency and precision, the Cas9 protein derived from the type II CRISPR system has been found to have applications in many fields of science. Although CRISPR–Cas9 allows easy genome editing and has a number of benefits, we should not ignore the important ethical and biosafety issues. Moreover, any tool that has great potential and offers significant capabilities carries a level of risk of being used for non-legal purposes. In this review, we present a brief history and mechanism of the CRISPR–Cas9 system. We also describe on the applications of this technology in gene regulation and genome editing; the treatment of cancer and other diseases; and limitations and concerns of the use of CRISPR–Cas9.
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Badhan, Sapna, Andrew S. Ball, and Nitin Mantri. "First Report of CRISPR/Cas9 Mediated DNA-Free Editing of 4CL and RVE7 Genes in Chickpea Protoplasts." International Journal of Molecular Sciences 22, no. 1 (January 1, 2021): 396. http://dx.doi.org/10.3390/ijms22010396.
Abstract:
The current genome editing system Clustered Regularly Interspaced Short Palindromic Repeats Cas9 (CRISPR/Cas9) has already confirmed its proficiency, adaptability, and simplicity in several plant-based applications. Together with the availability of a vast amount of genome data and transcriptome data, CRISPR/Cas9 presents a massive opportunity for plant breeders and researchers. The successful delivery of ribonucleoproteins (RNPs), which are composed of Cas9 enzyme and a synthetically designed single guide RNA (sgRNA) and are used in combination with various transformation methods or lately available novel nanoparticle-based delivery approaches, allows targeted mutagenesis in plants species. Even though this editing technique is limitless, it has still not been employed in many plant species to date. Chickpea is the second most crucial winter grain crop cultivated worldwide; there are currently no reports on CRISPR/Cas9 gene editing in chickpea. Here, we selected the 4-coumarate ligase (4CL) and Reveille 7 (RVE7) genes, both associated with drought tolerance for CRISPR/Cas9 editing in chickpea protoplast. The 4CL represents a key enzyme involved in phenylpropanoid metabolism in the lignin biosynthesis pathway. It regulates the accumulation of lignin under stress conditions in several plants. The RVE7 is a MYB transcription factor which is part of regulating circadian rhythm in plants. The knockout of these selected genes in the chickpea protoplast using DNA-free CRISPR/Cas9 editing represents a novel approach for achieving targeted mutagenesis in chickpea. Results showed high-efficiency editing was achieved for RVE7 gene in vivo compared to the 4CL gene. This study will help unravel the role of these genes under drought stress and understand the complex drought stress mechanism pathways. This is the first study in chickpea protoplast utilizing CRISPR/Cas9 DNA free gene editing of drought tolerance associated genes.
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Veillet, Florian, Laura Perrot, Anouchka Guyon-Debast, Marie-Paule Kermarrec, Laura Chauvin, Jean-Eric Chauvin, Jean-Luc Gallois, Marianne Mazier, and Fabien Nogué. "Expanding the CRISPR Toolbox in P. patens Using SpCas9-NG Variant and Application for Gene and Base Editing in Solanaceae Crops." International Journal of Molecular Sciences 21, no. 3 (February 4, 2020): 1024. http://dx.doi.org/10.3390/ijms21031024.
Abstract:
Genome editing has become a major tool for both functional studies and plant breeding in several species. Besides generating knockouts through the classical CRISPR-Cas9 system, recent development of CRISPR base editing holds great and exciting opportunities for the production of gain-of-function mutants. The PAM requirement is a strong limitation for CRISPR technologies such as base editing, because the base substitution mainly occurs in a small edition window. As precise single amino-acid substitution can be responsible for functions associated to some domains or agronomic traits, development of Cas9 variants with relaxed PAM recognition is of upmost importance for gene function analysis and plant breeding. Recently, the SpCas9-NG variant that recognizes the NGN PAM has been successfully tested in plants, mainly in monocotyledon species. In this work, we studied the efficiency of SpCas9-NG in the model moss Physcomitrella patens and two Solanaceae crops (Solanum lycopersicum and Solanum tuberosum) for both classical CRISPR-generated gene knock-out and cytosine base editing. We showed that the SpCas9-NG greatly expands the scope of genome editing by allowing the targeting of non-canonical NGT and NGA PAMs. The CRISPR toolbox developed in our study opens up new gene function analysis and plant breeding perspectives for model and crop plants.
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Vermersch, Eva, Charlène Jouve, and Jean-Sébastien Hulot. "CRISPR/Cas9 gene-editing strategies in cardiovascular cells." Cardiovascular Research 116, no. 5 (November 18, 2019): 894–907. http://dx.doi.org/10.1093/cvr/cvz250.
Abstract:
Abstract Cardiovascular diseases are among the main causes of morbidity and mortality in Western countries and considered as a leading public health issue. Therefore, there is a strong need for new disease models to support the development of novel therapeutics approaches. The successive improvement of genome editing tools with zinc finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs), and more recently with clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated 9 (Cas9) has enabled the generation of genetically modified cells and organisms with much greater efficiency and precision than before. The simplicity of CRISPR/Cas9 technology made it especially suited for different studies, both in vitro and in vivo, and has been used in multiple studies evaluating gene functions, disease modelling, transcriptional regulation, and testing of novel therapeutic approaches. Notably, with the parallel development of human induced pluripotent stem cells (hiPSCs), the generation of knock-out and knock-in human cell lines significantly increased our understanding of mutation impacts and physiopathological mechanisms within the cardiovascular domain. Here, we review the recent development of CRISPR–Cas9 genome editing, the alternative tools, the available strategies to conduct genome editing in cardiovascular cells with a focus on its use for correcting mutations in vitro and in vivo both in germ and somatic cells. We will also highlight that, despite its potential, CRISPR/Cas9 technology comes with important technical and ethical limitations. The development of CRISPR/Cas9 genome editing for cardiovascular diseases indeed requires to develop a specific strategy in order to optimize the design of the genome editing tools, the manipulation of DNA repair mechanisms, the packaging and delivery of the tools to the studied organism, and the assessment of their efficiency and safety.
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He, Ji, Riya Biswas, Piyush Bugde, Jiawei Li, Dong-Xu Liu, and Yan Li. "Application of CRISPR-Cas9 System to Study Biological Barriers to Drug Delivery." Pharmaceutics 14, no. 5 (April 20, 2022): 894. http://dx.doi.org/10.3390/pharmaceutics14050894.
Abstract:
In recent years, sequence-specific clustered regularly interspaced short palindromic repeats (CRISPR)-CRISPR-associated (Cas) systems have been widely used in genome editing of various cell types and organisms. The most developed and broadly used CRISPR-Cas system, CRISPR-Cas9, has benefited from the proof-of-principle studies for a better understanding of the function of genes associated with drug absorption and disposition. Genome-scale CRISPR-Cas9 knockout (KO) screen study also facilitates the identification of novel genes in which loss alters drug permeability across biological membranes and thus modulates the efficacy and safety of drugs. Compared with conventional heterogeneous expression models or other genome editing technologies, CRISPR-Cas9 gene manipulation techniques possess significant advantages, including ease of design, cost-effectiveness, greater on-target DNA cleavage activity and multiplexing capabilities, which makes it possible to study the interactions between membrane proteins and drugs more accurately and efficiently. However, many mechanistic questions and challenges regarding CRISPR-Cas9 gene editing are yet to be addressed, ranging from off-target effects to large-scale genetic alterations. In this review, an overview of the mechanisms of CRISPR-Cas9 in mammalian genome editing will be introduced, as well as the application of CRISPR-Cas9 in studying the barriers to drug delivery.
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Gong, Yan, Siyu Tian, Yang Xuan, and Shubiao Zhang. "Lipid and polymer mediated CRISPR/Cas9 gene editing." Journal of Materials Chemistry B 8, no. 20 (2020): 4369–86. http://dx.doi.org/10.1039/d0tb00207k.