Reprogramming Mycobacterium tuberculosis CRISPR System for Gene Editing and Genome-wide RNA Interference Screening
- Author:
Rahman KHAISTA
1
,
2
;
Jamal MUHAMMAD
;
Chen XI
;
Zhou WEI
;
Yang BIN
;
Zou YANYAN
;
Xu WEIZE
;
Lei YINGYING
;
Wu CHENGCHAO
;
Cao XIAOJIAN
;
Tyagi ROHIT
;
Naeem Ahsan MUHAMMAD
;
Lin DA
;
Habib ZESHAN
;
Peng NAN
;
F.Fu ZHEN
;
Cao GANG
Author Information
1. State Key Laboratory of Agricultural Microbiology,Huazhong Agricultural University,Wuhan 430070,China
2. College of Veterinary Medicine,Huazhong Agricultural University,Wuhan 430070,China
- Keywords:
Mycobacterium tuberculosis;
Type Ⅲ-A CRISPR system;
Gene editing;
Gene interference;
Genome-wide RNAi screening
- From:
Genomics, Proteomics & Bioinformatics
2022;(6):1180-1196
- CountryChina
- Language:Chinese
-
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 con-firmed 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.
