CRISPR Interference (CRISPRi) for Targeted Gene Silencing in Mycobacteria

Andrew I Wong; Jeremy M Rock

doi:10.1007/978-1-0716-1460-0_16

CRISPR Interference (CRISPRi) for Targeted Gene Silencing in Mycobacteria

Methods Mol Biol. 2021:2314:343-364. doi: 10.1007/978-1-0716-1460-0_16.

Authors

Andrew I Wong¹, Jeremy M Rock²

Affiliations

¹ Laboratory of Host-Pathogen Biology, The Rockefeller University, New York, NY, USA.
² Laboratory of Host-Pathogen Biology, The Rockefeller University, New York, NY, USA. rock@rockefeller.edu.

PMID: 34235662
DOI: 10.1007/978-1-0716-1460-0_16

Abstract

The genetic basis for Mycobacterium tuberculosis pathogenesis is incompletely understood. One reason for this knowledge gap is the relative difficulty of genetic manipulation of M. tuberculosis. To close this gap, we recently developed a robust CRISPR interference (CRISPRi) platform for programmable gene silencing in mycobacteria. In this chapter, we: (1) discuss some of the advantages and disadvantages of CRISPRi relative to more traditional genetic approaches; and (2) provide a protocol for the application of CRISPRi to reduce transcription of target genes in mycobacteria.

Keywords: CRISPR; CRISPR interference; CRISPRi; Gene knockdown; Gene silencing; Transcriptional silencing.

MeSH terms

Bacterial Proteins / antagonists & inhibitors*
Bacterial Proteins / genetics*
CRISPR-Cas Systems*
Gene Expression Regulation, Bacterial
Gene Silencing*
Genetic Engineering / methods*
Genetic Techniques
Mycobacterium tuberculosis / genetics*
Mycobacterium tuberculosis / growth & development

Substances

Bacterial Proteins