Journal
CELL CHEMICAL BIOLOGY
Volume 26, Issue 12, Pages 1732-+Publisher
CELL PRESS
DOI: 10.1016/j.chembiol.2019.09.003
Keywords
-
Categories
Funding
- National Natural Science Foundation of China [21922705, 91753127, 31700123]
- Shanghai Committee of Science and Technology, China [19QA1406000, 17ZR1449200]
- China Postdoctoral Science Foundation [2018M632190]
Ask authors/readers for more resources
The rapid emergence of extensively drug-resistant A. baumannii has posed a major threat to global public health, emphasizing the desperate need for novel therapeutic strategies. We report the development of a highly efficient genome-engineering platform in A. baumannii by coupling a Cas9 nuclease-mediated genome cleavage system with the RecAb recombination system. We applied the CRISPR-Cas9/RecAb system to dissect the oxidative stress-sensing mechanism of OxyR by performing alanine scanning mutagenesis of 13 residues residing in the H2O2-sensing pocket, pinpointing new vital factors for H2O2 sensing. Moreover, we developed a cytidine base-editing system, enabling programmed C to T conversions. Exploiting this powerful technique, we systematically investigated the drug-resistant mechanisms in a clinically isolated multidrug-resistant A. baumannii strain by generating premature stop codons in the possible resistance genes, unveiling distinct roles of these genes in drug resistance. The development of these genome-engineering methods will facilitate new therapeutic-means development in A. baumannii and related organisms.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available