清脆的
生物
计算生物学
基因组编辑
Cas9
合成生物学
单细胞分析
单元格排序
基因
遗传学
细胞
作者
Daniel Schraivogel,Lars M. Steinmetz,Leopold Parts
出处
期刊:Annual Review of Genetics
[Annual Reviews]
日期:2023-11-27
卷期号:57 (1): 223-244
被引量:5
标识
DOI:10.1146/annurev-genet-072920-013842
摘要
Assigning functions to genes and learning how to control their expression are part of the foundation of cell biology and therapeutic development. An efficient and unbiased method to accomplish this is genetic screening, which historically required laborious clone generation and phenotyping and is still limited by scale today. The rapid technological progress on modulating gene function with CRISPR-Cas and measuring it in individual cells has now relaxed the major experimental constraints and enabled pooled screening with complex readouts from single cells. Here, we review the principles and practical considerations for pooled single-cell CRISPR screening. We discuss perturbation strategies, experimental model systems, matching the perturbation to the individual cells, reading out cell phenotypes, and data analysis. Our focus is on single-cell RNA sequencing and cell sorting–based readouts, including image-enabled cell sorting. We expect this transformative approach to fuel biomedical research for the next several decades.
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