SILENCING OF COXIELLA BURNETII DOTB BASED ON CRISPR INTERFERENCE
Journal Title: Acta Parasitologica et Medica Entomologica Sinica - Year 2024, Vol 31, Issue 1
Abstract
[Objective] A recombinant plasmid for CRISPR interference (CRISPRi) that incorporates dCas9 from Streptococcus pyogenes was constructed to investigate the function of Coxiella burnetii genes through a prompt, simple method. [Methods] A single guide RNA (sgRNA) sequence targeting C. burnetii dotB was integrated into a recombinant plasmid, which was then transformed into C. burnetii using electroporation. Next, dCas9 expression was induced with dehydrated tetracycline (aTC) to inhibit the transcription of dotB. [Results] The results demonstrated that dCas9 can be expressed normally in the CRISPRi system through aTC induction. Suppression of dotB was observed at the transcriptional and translational levels. In addition, the intracellular reproduction of C. burnetii significantly decreased after the suppression of dotB expression. [Conclusion] Silencing technology for the C. burnetii gene based on CRISPRi was established in this study, providing support for studying the biological functions of C. burnetii genes.
Authors and Affiliations
Chun-yu ZHOU, Ming-liang ZHAO, Ru-xi CHENG, Shan ZHANG, Na-na LI, Yong-hui YU, Xuan OUYANG, Jun JIAO, Xiao-lu XIONG, Jia-ning ZHANG
Keywords
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- EP ID EP753966
- DOI 10-3969/j.issn.1005-0507-2024-01-002
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