清脆的
计算生物学
引导RNA
计算机科学
Cas9
回文
单细胞分析
基因组编辑
基因组学
生物
遗传学
细胞
基因组
基因
作者
Junyun Cheng,Gaole Lin,Tianhao Wang,Yunzhu Wang,Wenbo Guo,Jie Liao,Penghui Yang,Jie Chen,Xin Shao,Xiaoyan Lu,Ling Zhu,Yì Wáng,Xiaohui Fan
标识
DOI:10.1002/advs.202204484
摘要
Abstract 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.
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