Back to Search Start Over

Reprogramming Mycobacterium tuberculosis CRISPR System for Gene Editing and Genome-wide RNA Interference Screening.

Authors :
Rahman K
Jamal M
Chen X
Zhou W
Yang B
Zou Y
Xu W
Lei Y
Wu C
Cao X
Tyagi R
Naeem MA
Lin D
Habib Z
Peng N
Fu ZF
Cao G
Source :
Genomics, proteomics & bioinformatics [Genomics Proteomics Bioinformatics] 2022 Dec; Vol. 20 (6), pp. 1180-1196. Date of Electronic Publication: 2021 Dec 16.
Publication Year :
2022

Abstract

Mycobacterium tuberculosis is the causative agent of tuberculosis (TB), which is still the leading cause of mortality from a single infectious disease worldwide. The development of novel anti-TB drugs and vaccines is severely hampered by the complicated and time-consuming genetic manipulation techniques for M. tuberculosis. Here, we harnessed an endogenous type III-A CRISPR/Cas10 system of M. tuberculosis for efficient gene editing and RNA interference (RNAi). This simple and easy method only needs to transform a single mini-CRISPR array plasmid, thus avoiding the introduction of exogenous protein and minimizing proteotoxicity. We demonstrated that M. tuberculosis genes can be efficiently and specifically knocked in/out by this system as confirmed by DNA high-throughput sequencing. This system was further applied to single- and multiple-gene RNAi. Moreover, we successfully performed genome-wide RNAi screening to identify M. tuberculosis genes regulating in vitro and intracellular growth. This system can be extensively used for exploring the functional genomics of M. tuberculosis and facilitate the development of novel anti-TB drugs and vaccines.<br /> (Copyright © 2022. Published by Elsevier B.V.)

Details

Language :
English
ISSN :
2210-3244
Volume :
20
Issue :
6
Database :
MEDLINE
Journal :
Genomics, proteomics & bioinformatics
Publication Type :
Academic Journal
Accession number :
34923124
Full Text :
https://doi.org/10.1016/j.gpb.2021.01.008