Development and application of CRISPR assisted genome editing technology in Mycobacterium tuberculosis

doi:10.19982/j.issn.1000-6621.20220481

Abstract

Abstract:

Tuberculosis (TB), caused by Mycobacterium tuberculosis (MTB), remains a public health threat. To address this challenge, development of new anti-TB vaccines and drugs is urgent, which requires a more comprehensive understanding of gene function of MTB. CRISPR-Cas assisted genome editing has been widely used in prokaryotes and eukaryotes, whereas it was developed and applied in MTB until recently. Here, we reviewed recent advances in the development and the application of CRISPR assisted genome editing in MTB. This review particularly focused on the application of genome-wide CRISPR knockout and CRISPRi libraries for screening the genetic and chemical-genetic interaction profiling in MTB, aiming to deepen the understanding of gene editing in MTB and promote the basic research of MTB.

Key words: Mycobacterium tuberculosis, CRISPR-Cas system, Review literature as topic

CLC Number:

Qu Yunmo, Ding Xinyuan, Yan Meiyi, Guo Xiaopeng, Sun Yicheng. Development and application of CRISPR assisted genome editing technology in Mycobacterium tuberculosis[J]. Chinese Journal of Antituberculosis, 2023, 45(2): 208-214. doi: 10.19982/j.issn.1000-6621.20220481

Figures/Tables 2

References 30

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编辑类型	Cas种类	应用细菌	优点	缺点
CRISPR辅助的同源重组基因组编辑	CRISPR-Cas12a CRISPR-Cas9	耻垢分枝杆菌	提高细菌重组效率,不依赖抗性基因筛选突变菌株	目前无法应用于结核分枝杆菌
CRISPR辅助的非同源末端连接基因组编辑	CRISPR-Cas12a	耻垢分枝杆菌海分枝杆菌	快捷、高效,可应用于其他分枝杆菌的基因组编辑	无法应用于结核分枝杆菌
	CRISPR-Cas9_sth1	结核分枝杆菌耻垢分枝杆菌海分枝杆菌	快捷、高效,可应用于其他分枝杆菌的基因组编辑	只能用于基因敲除,无法实现基因点突变
CRISPR辅助的单碱基编辑技术	CRISPR-nCas9_sth1	耻垢分枝杆菌结核分枝杆菌	实现对分枝杆菌靶基因的点突变,可应用于其他分枝杆菌	只能用于特定位点的基因突变
CRISPRi调控的基因表达	CRISPR-dCas9_spy	耻垢分枝杆菌结核分枝杆菌	设计简单,易于改造,可应用于其他分枝杆菌	抑制基因表达的能力较弱,蛋白毒性大
	CRISPR-dCas9_sth1	耻垢分枝杆菌结核分枝杆菌	简单高效,高度抑制基因表达,且蛋白毒性小,有利于必需基因的研究。可应用于其他分枝杆菌	CRISPRi文库筛选对于下游基因具有极性效应