CRISPR interference and its applications

Seyedkhashayar Ghavami; Amir Pandi

doi:10.1016/bs.pmbts.2021.01.007

CRISPR interference and its applications

Prog Mol Biol Transl Sci. 2021:180:123-140. doi: 10.1016/bs.pmbts.2021.01.007. Epub 2021 Feb 12.

Authors

Seyedkhashayar Ghavami¹, Amir Pandi²

Affiliations

¹ Department of Biology, University of Groningen, Groningen, Netherlands.
² Department of Biochemistry and Synthetic Metabolism, Max-Planck Institute for Terrestrial Microbiology, Marburg, Germany. Electronic address: amir.pandi@mpi-marburg.mpg.de.

PMID: 33934834
DOI: 10.1016/bs.pmbts.2021.01.007

Abstract

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.

Keywords: CRISPR interference; CRISPR/dCas9; CRISPRi; Gene regulation; dCas9; sgRNA.

MeSH terms

RNA, Guide, CRISPR-Cas Systems* / genetics
Transcription Factors*

Substances

RNA, Guide, CRISPR-Cas Systems
Transcription Factors