Extensible combinatorial CRISPR screening in mammalian cells

The CRISPR-Cas system coupled with Combinatorial Genetics En Masse (CombiGEM) enables systematic analysis of high-order genetic perturbations that are important for understanding biological processes and discovering therapeutic target combinations. Here, we present detailed steps and technical consi...

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Detalles Bibliográficos
Autores principales: Zhou, Peng, Wan, Yuk Kei, Chan, Becky K.C., Choi, Gigi C.G., Wong, Alan S.L.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2021
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7809193/
https://www.ncbi.nlm.nih.gov/pubmed/33490975
http://dx.doi.org/10.1016/j.xpro.2020.100255
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author Zhou, Peng
Wan, Yuk Kei
Chan, Becky K.C.
Choi, Gigi C.G.
Wong, Alan S.L.
author_facet Zhou, Peng
Wan, Yuk Kei
Chan, Becky K.C.
Choi, Gigi C.G.
Wong, Alan S.L.
author_sort Zhou, Peng
collection PubMed
description The CRISPR-Cas system coupled with Combinatorial Genetics En Masse (CombiGEM) enables systematic analysis of high-order genetic perturbations that are important for understanding biological processes and discovering therapeutic target combinations. Here, we present detailed steps and technical considerations for building multiplexed guide RNA libraries and carrying out a combinatorial CRISPR screen in mammalian cells. We also present an analytical pipeline, CombiPIPE, for mapping two- and three-way genetic interactions. For complete details on the use and execution of this protocol, please refer to Zhou et al. (2020).
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spelling pubmed-78091932021-01-22 Extensible combinatorial CRISPR screening in mammalian cells Zhou, Peng Wan, Yuk Kei Chan, Becky K.C. Choi, Gigi C.G. Wong, Alan S.L. STAR Protoc Protocol The CRISPR-Cas system coupled with Combinatorial Genetics En Masse (CombiGEM) enables systematic analysis of high-order genetic perturbations that are important for understanding biological processes and discovering therapeutic target combinations. Here, we present detailed steps and technical considerations for building multiplexed guide RNA libraries and carrying out a combinatorial CRISPR screen in mammalian cells. We also present an analytical pipeline, CombiPIPE, for mapping two- and three-way genetic interactions. For complete details on the use and execution of this protocol, please refer to Zhou et al. (2020). Elsevier 2021-01-09 /pmc/articles/PMC7809193/ /pubmed/33490975 http://dx.doi.org/10.1016/j.xpro.2020.100255 Text en © 2020 The Author(s) http://creativecommons.org/licenses/by-nc-nd/4.0/ This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Protocol
Zhou, Peng
Wan, Yuk Kei
Chan, Becky K.C.
Choi, Gigi C.G.
Wong, Alan S.L.
Extensible combinatorial CRISPR screening in mammalian cells
title Extensible combinatorial CRISPR screening in mammalian cells
title_full Extensible combinatorial CRISPR screening in mammalian cells
title_fullStr Extensible combinatorial CRISPR screening in mammalian cells
title_full_unstemmed Extensible combinatorial CRISPR screening in mammalian cells
title_short Extensible combinatorial CRISPR screening in mammalian cells
title_sort extensible combinatorial crispr screening in mammalian cells
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7809193/
https://www.ncbi.nlm.nih.gov/pubmed/33490975
http://dx.doi.org/10.1016/j.xpro.2020.100255
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