SILENCING OF COXIELLA BURNETII DOTB BASED ON CRISPR INTERFERENCE
10.3969/j.issn.1005-0507.2024.01.002
- VernacularTitle:基于CRISPRi的贝氏柯克斯体dotB基因沉默
- Author:
Chun-Yu ZHOU
1
;
Ming-Liang ZHAO
;
Ru-Xi CHENG
;
Shan ZHANG
;
Na-Na LI
;
Yong-Hui YU
;
Xuan OUYANG
;
Jun JIAO
;
Xiao-Lu XIONG
;
Jia-Ning ZHANG
Author Information
1. 牡丹江医学院公共卫生学院, 黑龙江牡丹江 157000;军事科学院军事医学研究院微生物流行病研究所, 病原微生物生物安全全国重点实验室, 北京 100071
- Keywords:
Coxiella burnetii;
Silencing;
CRISPRi;
dCas9;
DotB
- From:
Acta Parasitologica et Medica Entomologica Sinica
2024;31(1):6-11
- CountryChina
- Language:Chinese
-
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.
