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Academic literature on the topic 'FUCCI reporters'

Author: Grafiati
Published: 10 December 2022
Last updated: 28 January 2023

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Journal articles on the topic "FUCCI reporters"

1

Singh, Amar M., Robert Trost, Benjamin Boward, and Stephen Dalton. "Utilizing FUCCI reporters to understand pluripotent stem cell biology." Methods 101 (May 2016): 4–10. http://dx.doi.org/10.1016/j.ymeth.2015.09.020.

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Yano, Shuya, Hiroshi Tazawa, Shunsuke Kagawa, Toshiyoshi Fujiwara, and Robert M. Hoffman. "FUCCI Real-Time Cell-Cycle Imaging as a Guide for Designing Improved Cancer Therapy: A Review of Innovative Strategies to Target Quiescent Chemo-Resistant Cancer Cells." Cancers 12, no. 9 (September 17, 2020): 2655. http://dx.doi.org/10.3390/cancers12092655.

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Progress in chemotherapy of solid cancer has been tragically slow due, in large part, to the chemoresistance of quiescent cancer cells in tumors. The fluorescence ubiquitination cell-cycle indicator (FUCCI) was developed in 2008 by Miyawaki et al., which color-codes the phases of the cell cycle in real-time. FUCCI utilizes genes linked to different color fluorescent reporters that are only expressed in specific phases of the cell cycle and can, thereby, image the phases of the cell cycle in real-time. Intravital real-time FUCCI imaging within tumors has demonstrated that an established tumor comprises a majority of quiescent cancer cells and a minor population of cycling cancer cells located at the tumor surface or in proximity to tumor blood vessels. In contrast to most cycling cancer cells, quiescent cancer cells are resistant to cytotoxic chemotherapy, most of which target cells in S/G2/M phases. The quiescent cancer cells can re-enter the cell cycle after surviving treatment, which suggests the reason why most cytotoxic chemotherapy is often ineffective for solid cancers. Thus, quiescent cancer cells are a major impediment to effective cancer therapy. FUCCI imaging can be used to effectively target quiescent cancer cells within tumors. For example, we review how FUCCI imaging can help to identify cell-cycle-specific therapeutics that comprise decoy of quiescent cancer cells from G1 phase to cycling phases, trapping the cancer cells in S/G2 phase where cancer cells are mostly sensitive to cytotoxic chemotherapy and eradicating the cancer cells with cytotoxic chemotherapy most active against S/G2 phase cells. FUCCI can readily image cell-cycle dynamics at the single cell level in real-time in vitro and in vivo. Therefore, visualizing cell cycle dynamics within tumors with FUCCI can provide a guide for many strategies to improve cell-cycle targeting therapy for solid cancers.
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Zheng, Lixia, Zihao Wang, Jianyong Du, Xiaojun Zhu, and Jing-Wei Xiong. "Protocol to identify small molecules promoting rat and mouse cardiomyocyte proliferation based on the FUCCI and MADM reporters." STAR Protocols 3, no. 4 (December 2022): 101903. http://dx.doi.org/10.1016/j.xpro.2022.101903.

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Ippati, Stefania, Yuanyuan Deng, Julia van der Hoven, Celine Heu, Annika van Hummel, Sook Wern Chua, Esmeralda Paric, et al. "Rapid initiation of cell cycle reentry processes protects neurons from amyloid-β toxicity." Proceedings of the National Academy of Sciences 118, no. 12 (March 18, 2021): e2011876118. http://dx.doi.org/10.1073/pnas.2011876118.

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Neurons are postmitotic cells. Reactivation of the cell cycle by neurons has been reported in Alzheimer’s disease (AD) brains and models. This gave rise to the hypothesis that reentering the cell cycle renders neurons vulnerable and thus contributes to AD pathogenesis. Here, we use the fluorescent ubiquitination-based cell cycle indicator (FUCCI) technology to monitor the cell cycle in live neurons. We found transient, self-limited cell cycle reentry activity in naive neurons, suggesting that their postmitotic state is a dynamic process. Furthermore, we observed a diverse response to oligomeric amyloid-β (oAβ) challenge; neurons without cell cycle reentry activity would undergo cell death without activating the FUCCI reporter, while neurons undergoing cell cycle reentry activity at the time of the oAβ challenge could maintain and increase FUCCI reporter signal and evade cell death. Accordingly, we observed marked neuronal FUCCI positivity in the brains of human mutant Aβ precursor protein transgenic (APP23) mice together with increased neuronal expression of the endogenous cell cycle control protein geminin in the brains of 3-mo-old APP23 mice and human AD brains. Taken together, our data challenge the current view on cell cycle in neurons and AD, suggesting that pathways active during early cell cycle reentry in neurons protect from Aβ toxicity.
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Manolopoulou, Marika, Brittany K. Matlock, Stellor Nlandu-Khodo, Alan J. Simmons, Ken S. Lau, Melanie Phillips-Mignemi, Alla Ivanova, Catherine E. Alford, David K. Flaherty, and Leslie S. Gewin. "Novel kidney dissociation protocol and image-based flow cytometry facilitate improved analysis of injured proximal tubules." American Journal of Physiology-Renal Physiology 316, no. 5 (May 1, 2019): F847—F855. http://dx.doi.org/10.1152/ajprenal.00354.2018.

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Flow cytometry studies on injured kidney tubules are complicated by the low yield of nucleated single cells. Furthermore, cell-specific responses such as cell cycle dynamics in vivo have conventionally relied on indirect immunohistochemistry and proximal tubule markers that may be downregulated in injury. Here, we report a new tissue dissociation protocol for the kidney with an early fixation step that greatly enhances the yield of single cells. Genetic labeling of the proximal tubule with either mT/mG “tomato” or R26Fucci2aR (Fucci) cell cycle reporter mice allows us to follow proximal tubule-specific changes in cell cycle after renal injury. Image-based flow cytometry (FlowSight) enables gating of the cell cycle and concurrent visualization of the cells with bright field and fluorescence. We used the Fucci mouse in conjunction with FlowSight to identify a discrete polyploid population in proximal tubules after aristolochic acid injury. The tissue dissociation protocol in conjunction with genetic labeling and image-based flow cytometry is a tool that can improve our understanding of any discrete cell population after injury.
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Takahashi, Kei, Ryo Tanabe, Shogo Ehata, Shimpei I. Kubota, Yasuyuki Morishita, Hiroki R. Ueda, and Kohei Miyazono. "Visualization of the cancer cell cycle by tissue‐clearing technology using the Fucci reporter system." Cancer Science 112, no. 9 (July 7, 2021): 3796–809. http://dx.doi.org/10.1111/cas.15034.

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7

Zambon, Alexander C., Tom Hsu, Seunghee Erin Kim, Miranda Klinck, Jennifer Stowe, Lindsay M. Henderson, Donald Singer, et al. "Methods and sensors for functional genomic studies of cell-cycle transitions in single cells." Physiological Genomics 52, no. 10 (October 1, 2020): 468–77. http://dx.doi.org/10.1152/physiolgenomics.00065.2020.

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Much of our understanding of the regulatory mechanisms governing the cell cycle in mammals has relied heavily on methods that measure the aggregate state of a population of cells. While instrumental in shaping our current understanding of cell proliferation, these approaches mask the genetic signatures of rare subpopulations such as quiescent (G0) and very slowly dividing (SD) cells. Results described in this study and those of others using single-cell analysis reveal that even in clonally derived immortalized cancer cells, ∼1–5% of cells can exhibit G0 and SD phenotypes. Therefore to enable the study of these rare cell phenotypes we established an integrated molecular, computational, and imaging approach to track, isolate, and genetically perturb single cells as they proliferate. A genetically encoded cell-cycle reporter (K67p-FUCCI) was used to track single cells as they traversed the cell cycle. A set of R-scripts were written to quantify K67p-FUCCI over time. To enable the further study G0 and SD phenotypes, we retrofitted a live cell imaging system with a micromanipulator to enable single-cell targeting for functional validation studies. Single-cell analysis revealed HT1080 and MCF7 cells had a doubling time of ∼24 and ∼48 h, respectively, with high duration variability in G1 and G2 phases. Direct single-cell microinjection of mRNA encoding (GFP) achieves detectable GFP fluorescence within ∼5 h in both cell types. These findings coupled with the possibility of targeting several hundreds of single cells improves throughput and sensitivity over conventional methods to study rare cell subpopulations.
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Mubarok, Wildan, Kelum Chamara Manoj Lakmal Elvitigala, Masaki Nakahata, Masaru Kojima, and Shinji Sakai. "Modulation of Cell-Cycle Progression by Hydrogen Peroxide-Mediated Cross-Linking and Degradation of Cell-Adhesive Hydrogels." Cells 11, no. 5 (March 3, 2022): 881. http://dx.doi.org/10.3390/cells11050881.

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The cell cycle is known to be regulated by features such as the mechanical properties of the surrounding environment and interaction of cells with the adhering substrates. Here, we investigated the possibility of regulating cell-cycle progression of the cells on gelatin/hyaluronic acid composite hydrogels obtained through hydrogen peroxide (H2O2)-mediated cross-linking and degradation of the polymers by varying the exposure time to H2O2 contained in the air. The stiffness of the hydrogel varied with the exposure time. Human cervical cancer cells (HeLa) and mouse mammary gland epithelial cells (NMuMG) expressing cell-cycle reporter Fucci2 showed the exposure-time-dependent different cell-cycle progressions on the hydrogels. Although HeLa/Fucci2 cells cultured on the soft hydrogel (Young’s modulus: 0.20 and 0.40 kPa) obtained through 15 min and 120 min of the H2O2 exposure showed a G2/M-phase arrest, NMuMG cells showed a G1-phase arrest. Additionally, the cell-cycle progression of NMuMG cells was not only governed by the hydrogel stiffness, but also by the low-molecular-weight HA resulting from H2O2-mediated degradation. These results indicate that H2O2-mediated cross-linking and degradation of gelatin/hyaluronic acid composite hydrogel could be used to control the cell adhesion and cell-cycle progression.
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Yano, Shuya, Hiroshi Tazawa, Hiroyuki Kishimoto, Shunsuke Kagawa, Toshiyoshi Fujiwara, and Robert M. Hoffman. "Real-Time Fluorescence Image-Guided Oncolytic Virotherapy for Precise Cancer Treatment." International Journal of Molecular Sciences 22, no. 2 (January 17, 2021): 879. http://dx.doi.org/10.3390/ijms22020879.

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Oncolytic virotherapy is one of the most promising, emerging cancer therapeutics. We generated three types of telomerase-specific replication-competent oncolytic adenovirus: OBP-301; a green fluorescent protein (GFP)-expressing adenovirus, OBP-401; and Killer-Red-armed OBP-301. These oncolytic adenoviruses are driven by the human telomerase reverse transcriptase (hTERT) promoter; therefore, they conditionally replicate preferentially in cancer cells. Fluorescence imaging enables visualization of invasion and metastasis in vivo at the subcellular level; including molecular dynamics of cancer cells, resulting in greater precision therapy. In the present review, we focused on fluorescence imaging applications to develop precision targeting for oncolytic virotherapy. Cell-cycle imaging with the fluorescence ubiquitination cell cycle indicator (FUCCI) demonstrated that combination therapy of an oncolytic adenovirus and a cytotoxic agent could precisely target quiescent, chemoresistant cancer stem cells (CSCs) based on decoying the cancer cells to cycle to S-phase by viral treatment, thereby rendering them chemosensitive. Non-invasive fluorescence imaging demonstrated that complete tumor resection with a precise margin, preservation of function, and prevention of distant metastasis, was achieved with fluorescence-guided surgery (FGS) with a GFP-reporter adenovirus. A combination of fluorescence imaging and laser ablation using a KillerRed-protein reporter adenovirus resulted in effective photodynamic cancer therapy (PDT). Thus, imaging technology and the designer oncolytic adenoviruses may have clinical potential for precise cancer targeting by indicating the optimal time for administering therapeutic agents; accurate surgical guidance for complete resection of tumors; and precise targeted cancer-specific photosensitization.
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Abe, T., A. Sakaue-Sawano, H. Kiyonari, G. Shioi, K. i. Inoue, T. Horiuchi, K. Nakao, A. Miyawaki, S. Aizawa, and T. Fujimori. "Visualization of cell cycle in mouse embryos with Fucci2 reporter directed by Rosa26 promoter." Development 140, no. 1 (November 22, 2012): 237–46. http://dx.doi.org/10.1242/dev.084111.

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Book chapters on the topic "FUCCI reporters"

1

Chappell, James, Ben Boward, and Stephen Dalton. "Expanding the Utility of FUCCI Reporters Using FACS-Based Omics Analysis." In Embryonic Stem Cell Protocols, 101–10. New York, NY: Springer New York, 2015. http://dx.doi.org/10.1007/7651_2015_214.

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Conference papers on the topic "FUCCI reporters"

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Pineda, Gabriel, Florence Lambert-Fliszar, Gennarina L. Riso, Kathleen M. Kane, and Catriona Jamieson. "Abstract LB-159: Characterization of cell-cycle progression using a novel bi-cistronic lentiviral FUCCI reporter." In Proceedings: AACR Annual Meeting 2014; April 5-9, 2014; San Diego, CA. American Association for Cancer Research, 2014. http://dx.doi.org/10.1158/1538-7445.am2014-lb-159.

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