Modulating Pathogenesis with Mobile-CRISPRi

Qu, Jiuxin1,2,4; Prasad, Neha K.1,2; Yu, Michelle A.2; Chen, Shuyan1,2,5; Lyden, Amy1; Herrera, Nadia1,2; Silvis, Melanie R.3; Crawford, Emily1,3; Looney, Mark R.2; Peters, Jason M.3,6; Rosenberg, Oren S.1,2

2019-11

10.1128/JB.00304-19

JOURNAL OF BACTERIOLOGY   影响因子和分区

0021-9193

1098-5530

Conditionally essential (CE) genes are required by pathogenic bacteria to establish and maintain infections. CE genes encode virulence factors, such as secretion systems and effector proteins, as well as biosynthetic enzymes that produce metabolites not found in the host environment. Due to their outsized importance in pathogenesis, CE gene products are attractive targets for the next generation of antimicrobials. However, the precise manipulation of CE gene expression in the context of infection is technically challenging, limiting our ability to understand the roles of CE genes in pathogenesis and accordingly design effective inhibitors. We previously developed a suite of CRISPR interference-based gene knockdown tools that are transferred by conjugation and stably integrate into bacterial genomes that we call Mobile-CRISPRi. Here, we show the efficacy of Mobile-CRISPRi in controlling CE gene expression in an animal infection model. We optimize Mobile-CRISPRi in Pseudomonas aeruginosa for use in a murine model of pneumonia by tuning the expression of CRISPRi components to avoid nonspecific toxicity. As a proof of principle, we demonstrate that knock down of a CE gene encoding the type III secretion system (T3SS) activator ExsA blocks effector protein secretion in culture and attenuates virulence in mice. We anticipate that Mobile-CRISPRi will be a valuable tool to probe the function of CE genes across many bacterial species and pathogenesis models.

CRISPRi conditionally essential genes type III secretion Pseudomonas aeruginosa infection model virulence lifestyle genes

SCI

英语

其他

Special Support Fund of Shenzhen for Introduced High-Level Medical Team and Translational medicine of Biochip in clinical laboratory[SZSM201412005] ; Special Support Fund of Shenzhen for Introduced High-Level Medical Team and Translational medicine of Biochip in clinical laboratory[5R01EB024014]

Microbiology

Microbiology

WOS:000491960200001

AMER SOC MICROBIOLOGY

MICROBIOLOGY

Web of Science

1.Chan Zuckerberg Biohub, San Francisco, CA 94158 USA
2.Univ Calif San Francisco, Dept Med, San Francisco, CA 94143 USA
3.Univ Calif San Francisco, Dept Microbiol & Immunol, San Francisco, CA 94143 USA
4.South Univ Sci & Technol China, Third Peoples Hosp Shenzhen, Dept Clin Lab, Shenzhen, Guangdong, Peoples R China
5.South Univ Sci & Technol China, Third Peoples Hosp Shenzhen, Dept Clin Res, Shenzhen, Guangdong, Peoples R China
6.Univ Wisconsin, Div Pharmaceut Sci, Dept Bacteriol, Great Lakes Bioenergy Res Ctr,Dept Med Microbiol, Madison, WI 53706 USA

Qu, Jiuxin,Prasad, Neha K.,Yu, Michelle A.,et al. Modulating Pathogenesis with Mobile-CRISPRi[J]. JOURNAL OF BACTERIOLOGY,2019,201(22).

Qu, Jiuxin.,Prasad, Neha K..,Yu, Michelle A..,Chen, Shuyan.,Lyden, Amy.,...&Rosenberg, Oren S..(2019).Modulating Pathogenesis with Mobile-CRISPRi.JOURNAL OF BACTERIOLOGY,201(22).

Qu, Jiuxin,et al."Modulating Pathogenesis with Mobile-CRISPRi".JOURNAL OF BACTERIOLOGY 201.22(2019).