Programmable Base Editing in Mycobacterium tuberculosis Using an Engineered CRISPR RNA-Guided Cytidine Deaminase

Ding，Xin Yuan1; Li，Si Shang1; Geng，Yi Man2; Yan，Mei Yi1; Li，Guo Bao3; Zhang，Guo Liang3; Sun，Yi Cheng1

2021

10.3389/fgeed.2021.734436

Frontiers in Genome Editing   影响因子和分区

2673-3439

Multidrug-resistant Mycobacterium tuberculosis (Mtb) infection seriously endangers global human health, creating an urgent need for new treatment strategies. Efficient genome editing tools can facilitate identification of key genes and pathways involved in bacterial physiology, pathogenesis, and drug resistance mechanisms, and thus contribute to the development of novel treatments for drug-resistant tuberculosis. Here, we report a two-plasmid system, MtbCBE, used to inactivate genes and introduce point mutations in Mtb. In this system, the assistant plasmid pRecX-NucS expresses RecX and NucS to repress RecA-dependent and NucS-dependent DNA repair systems, and the base editor plasmid pCBE expresses a fusion protein combining cytidine deaminase APOBEC1, Cas9 nickase (nCas9), and uracil DNA glycosylase inhibitor (UGI). Together, the two plasmids enabled efficient G:C to A:T base pair conversion at desired sites in the Mtb genome. The successful development of a base editing system will facilitate elucidation of the molecular mechanisms underlying Mtb pathogenesis and drug resistance and provide critical inspiration for the development of base editing tools in other microbes.

base editing CRISPR cytidine deaminase genome editing mycobacteria

英语

其他

2-s2.0-85128918160

Scopus

1.NHC Key Laboratory of Systems Biology of Pathogens,Institute of Pathogen Biology,Center for Tuberculosis Research,Chinese Academy of Medical Sciences and Peking Union Medical College,Beijing,China
2.Department of Clinical Laboratory,Henan Provincial People’s Hospital,People’s Hospital of Zhengzhou University,Zhengzhou,China
3.National Clinical Research Center for Infectious Diseases,Shenzhen Third People’s Hospital,Southern University of Science and Technology,Shenzhen,China

Ding，Xin Yuan,Li，Si Shang,Geng，Yi Man,et al. Programmable Base Editing in Mycobacterium tuberculosis Using an Engineered CRISPR RNA-Guided Cytidine Deaminase[J]. Frontiers in Genome Editing,2021,3.

Ding，Xin Yuan.,Li，Si Shang.,Geng，Yi Man.,Yan，Mei Yi.,Li，Guo Bao.,...&Sun，Yi Cheng.(2021).Programmable Base Editing in Mycobacterium tuberculosis Using an Engineered CRISPR RNA-Guided Cytidine Deaminase.Frontiers in Genome Editing,3.

Ding，Xin Yuan,et al."Programmable Base Editing in Mycobacterium tuberculosis Using an Engineered CRISPR RNA-Guided Cytidine Deaminase".Frontiers in Genome Editing 3(2021).