Department of Biology, University of Groningen, Groningen, Netherlands.
Department of Biochemistry and Synthetic Metabolism, Max-Planck Institute for Terrestrial Microbiology, Marburg, Germany.
Prog Mol Biol Transl Sci. 2021;180:123-140. doi: 10.1016/bs.pmbts.2021.01.007. Epub 2021 Feb 12.
Sequence-specific control of gene expression is a powerful tool for identifying and studying gene functions and cellular processes. CRISPR interference (CRISPRi) is an RNA-based method for highly specific silencing of the transcription in prokaryotic or eukaryotic cells. The typical CRISPRi system is a type II CRISPR (clustered regularly interspaced palindromic repeats) machinery of Streptococcus pyogenes. CRISPRi requires two main components: A catalytically inactivated Cas9, namely dCas9 and a guide RNA (sgRNA). These two components associate and form a DNA recognition complex. The dCas9/sgRNA complex then specifically binds to the target DNA complementary with the sgRNA and sterically prevents the association of the promoter or transcription factors with their trans-acting sequences or blocks the transcription elongation. This chapter discusses CRISPRi structure, mechanism and its applications.
序列特异性基因表达调控是鉴定和研究基因功能和细胞过程的有力工具。CRISPR 干扰（CRISPRi）是一种基于 RNA 的方法，可高度特异性地抑制原核或真核细胞中的转录。典型的 CRISPRi 系统是来自酿脓链球菌的 II 型 CRISPR（成簇的规律间隔的短回文重复序列）机制。CRISPRi 需要两个主要组件：一种无催化活性的 Cas9，即 dCas9 和向导 RNA（sgRNA）。这两个组件结合形成 DNA 识别复合物。然后，dCas9/sgRNA 复合物特异性地与与 sgRNA 互补的靶 DNA 结合，并通过空间位阻防止启动子或转录因子与其反式作用序列结合或阻断转录延伸。本章讨论了 CRISPRi 的结构、机制及其应用。
