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Massively Parallel CRISPR‐Based Genetic Perturbation Screening at Single‐Cell Resolution
The clustered regularly interspaced short palindromic repeats (CRISPR)‐based genetic screening has been demonstrated as a powerful approach for unbiased functional genomics research. Single‐cell CRISPR screening (scCRISPR) techniques, which result from the combination of single‐cell toolkits and CRI...
Autores principales: | , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
John Wiley and Sons Inc.
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9896079/ https://www.ncbi.nlm.nih.gov/pubmed/36504444 http://dx.doi.org/10.1002/advs.202204484 |
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author | Cheng, Junyun Lin, Gaole Wang, Tianhao Wang, Yunzhu Guo, Wenbo Liao, Jie Yang, Penghui Chen, Jie Shao, Xin Lu, Xiaoyan Zhu, Ling Wang, Yi Fan, Xiaohui |
author_facet | Cheng, Junyun Lin, Gaole Wang, Tianhao Wang, Yunzhu Guo, Wenbo Liao, Jie Yang, Penghui Chen, Jie Shao, Xin Lu, Xiaoyan Zhu, Ling Wang, Yi Fan, Xiaohui |
author_sort | Cheng, Junyun |
collection | PubMed |
description | The clustered regularly interspaced short palindromic repeats (CRISPR)‐based genetic screening has been demonstrated as a powerful approach for unbiased functional genomics research. Single‐cell CRISPR screening (scCRISPR) techniques, which result from the combination of single‐cell toolkits and CRISPR screening, allow dissecting regulatory networks in complex biological systems at unprecedented resolution. These methods allow cells to be perturbed en masse using a pooled CRISPR library, followed by high‐content phenotyping. This is technically accomplished by annotating cells with sgRNA‐specific barcodes or directly detectable sgRNAs. According to the integration of distinct single‐cell technologies, these methods principally fall into four categories: scCRISPR with RNA‐seq, scCRISPR with ATAC‐seq, scCRISPR with proteome probing, and imaging‐based scCRISPR. scCRISPR has deciphered genotype–phenotype relationships, genetic regulations, tumor biological issues, and neuropathological mechanisms. This review provides insight into the technical breakthrough of scCRISPR by systematically summarizing the advancements of various scCRISPR methodologies and analyzing their merits and limitations. In addition, an application‐oriented approach guide is offered to meet researchers’ individualized demands. |
format | Online Article Text |
id | pubmed-9896079 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | John Wiley and Sons Inc. |
record_format | MEDLINE/PubMed |
spelling | pubmed-98960792023-02-08 Massively Parallel CRISPR‐Based Genetic Perturbation Screening at Single‐Cell Resolution Cheng, Junyun Lin, Gaole Wang, Tianhao Wang, Yunzhu Guo, Wenbo Liao, Jie Yang, Penghui Chen, Jie Shao, Xin Lu, Xiaoyan Zhu, Ling Wang, Yi Fan, Xiaohui Adv Sci (Weinh) Reviews The clustered regularly interspaced short palindromic repeats (CRISPR)‐based genetic screening has been demonstrated as a powerful approach for unbiased functional genomics research. Single‐cell CRISPR screening (scCRISPR) techniques, which result from the combination of single‐cell toolkits and CRISPR screening, allow dissecting regulatory networks in complex biological systems at unprecedented resolution. These methods allow cells to be perturbed en masse using a pooled CRISPR library, followed by high‐content phenotyping. This is technically accomplished by annotating cells with sgRNA‐specific barcodes or directly detectable sgRNAs. According to the integration of distinct single‐cell technologies, these methods principally fall into four categories: scCRISPR with RNA‐seq, scCRISPR with ATAC‐seq, scCRISPR with proteome probing, and imaging‐based scCRISPR. scCRISPR has deciphered genotype–phenotype relationships, genetic regulations, tumor biological issues, and neuropathological mechanisms. This review provides insight into the technical breakthrough of scCRISPR by systematically summarizing the advancements of various scCRISPR methodologies and analyzing their merits and limitations. In addition, an application‐oriented approach guide is offered to meet researchers’ individualized demands. John Wiley and Sons Inc. 2022-12-11 /pmc/articles/PMC9896079/ /pubmed/36504444 http://dx.doi.org/10.1002/advs.202204484 Text en © 2022 The Authors. Advanced Science published by Wiley‐VCH GmbH https://creativecommons.org/licenses/by/4.0/This is an open access article under the terms of the http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Reviews Cheng, Junyun Lin, Gaole Wang, Tianhao Wang, Yunzhu Guo, Wenbo Liao, Jie Yang, Penghui Chen, Jie Shao, Xin Lu, Xiaoyan Zhu, Ling Wang, Yi Fan, Xiaohui Massively Parallel CRISPR‐Based Genetic Perturbation Screening at Single‐Cell Resolution |
title | Massively Parallel CRISPR‐Based Genetic Perturbation Screening at Single‐Cell Resolution |
title_full | Massively Parallel CRISPR‐Based Genetic Perturbation Screening at Single‐Cell Resolution |
title_fullStr | Massively Parallel CRISPR‐Based Genetic Perturbation Screening at Single‐Cell Resolution |
title_full_unstemmed | Massively Parallel CRISPR‐Based Genetic Perturbation Screening at Single‐Cell Resolution |
title_short | Massively Parallel CRISPR‐Based Genetic Perturbation Screening at Single‐Cell Resolution |
title_sort | massively parallel crispr‐based genetic perturbation screening at single‐cell resolution |
topic | Reviews |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9896079/ https://www.ncbi.nlm.nih.gov/pubmed/36504444 http://dx.doi.org/10.1002/advs.202204484 |
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