Shanxi Provincial Key Laboratory for Medical Molecular Cell Biology, Key Laboratory of Chemical Biology and Molecular Engineering of Ministry of Education and Institute of Biomedical Sciences, Shanxi University, Taiyuan 030006, China.
Int J Mol Sci. 2024 Jan 4;25(1):658. doi: 10.3390/ijms25010658.
High-throughput genetic screening is useful for discovering critical genes or gene sequences that trigger specific cell functions and/or phenotypes. Loss-of-function genetic screening is mainly achieved through RNA interference (RNAi), CRISPR knock-out (CRISPRko), and CRISPR interference (CRISPRi) technologies. Gain-of-function genetic screening mainly depends on the overexpression of a cDNA library and CRISPR activation (CRISPRa). Base editing can perform both gain- and loss-of-function genetic screening. This review discusses genetic screening techniques based on Cas9 nuclease, including Cas9-mediated genome knock-out and dCas9-based gene activation and interference. We compare these methods with previous genetic screening techniques based on RNAi and cDNA library overexpression and propose future prospects and applications for CRISPR screening.
高通量基因筛选对于发现触发特定细胞功能和/或表型的关键基因或基因序列非常有用。失活基因筛选主要通过 RNA 干扰（RNAi）、CRISPR 敲除（CRISPRko）和 CRISPR 干扰（CRISPRi）技术来实现。功能获得性基因筛选主要依赖于 cDNA 文库的过表达和 CRISPR 激活（CRISPRa）。碱基编辑可以进行功能获得和功能丧失的基因筛选。本文综述了基于 Cas9 核酸酶的基因筛选技术，包括 Cas9 介导的基因组敲除和 dCas9 介导的基因激活和干扰。我们将这些方法与基于 RNAi 和 cDNA 文库过表达的先前基因筛选技术进行了比较，并对 CRISPR 筛选的未来前景和应用进行了展望。
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