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Abstract：
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.
摘要：
高通量遗传筛选可用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。功能缺失遗传筛选主要通过RNA干扰(RNAi)实现,CRISPR敲除（CRISPRko），和CRISPR干扰(CRISPRI)技术。功能获得遗传筛选主要取决于cDNA文库的过表达和CRISPR激活(CRISPRa)。碱基编辑可以进行功能增益和功能丧失的遗传筛选。本文综述了基于Cas9核酸酶的遗传筛选技术，包括Cas9介导的基因组敲除和基于dCas9的基因激活和干扰。我们将这些方法与以前基于RNAi和cDNA文库过表达的遗传筛选技术进行了比较，并提出了CRISPR筛选的未来前景和应用。
