| Autor: |
Rahman, Khaista, Jamal, Muhammad, Chen, Xi, Zhou, Wei, Yang, Bin, Zou, Yanyan, Xu, Weize, Lei, Yingying, Wu, Chengchao, Cao, Xiaojian, Tyagi, Rohit, Naeem, Muhammad Ahsan, Lin, Da, Habib, Zeshan, Peng, Nan, Fu, Zhen F., Cao, Gang |
| Zdroj: |
Genomics Proteomics and Bioinformatics; 20230101, Issue: Preprints |
| Abstrakt: |
Mycobacterium tuberculosisis 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. tuberculosisfor efficient gene editingand 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. tuberculosisgenes 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 screeningto identify M. tuberculosisgenes regulating in vitroand intracellular growth. This system can be extensively used for exploring the functional genomics of M. tuberculosisand facilitate the development of novel anti-TB drugs and vaccines. |
| Databáze: |
Supplemental Index |
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