Exploration of Bacterial Bottlenecks and Streptococcus pneumoniae Pathogenesis by CRISPRi-Seq.

Details

Serval ID
serval:BIB_E58DFC96E446
Type
Article: article from journal or magazin.
Collection
Publications
Institution
Title
Exploration of Bacterial Bottlenecks and Streptococcus pneumoniae Pathogenesis by CRISPRi-Seq.
Journal
Cell host & microbe
Author(s)
Liu X., Kimmey J.M., Matarazzo L., de Bakker V., Van Maele L., Sirard J.C., Nizet V., Veening J.W.
ISSN
1934-6069 (Electronic)
ISSN-L
1931-3128
Publication state
Published
Issued date
13/01/2021
Peer-reviewed
Oui
Volume
29
Number
1
Pages
107-120.e6
Language
english
Notes
Publication types: Journal Article
Publication Status: ppublish
Abstract
Streptococcus pneumoniae is an opportunistic human pathogen that causes invasive diseases, including pneumonia, with greater health risks upon influenza A virus (IAV) co-infection. To facilitate pathogenesis studies in vivo, we developed an inducible CRISPR interference system that enables genome-wide fitness testing in one sequencing step (CRISPRi-seq). We applied CRISPRi-seq to assess bottlenecks and identify pneumococcal genes important in a murine pneumonia model. A critical bottleneck occurs at 48 h with few bacteria causing systemic infection. This bottleneck is not present during IAV superinfection, facilitating identification of pneumococcal pathogenesis-related genes. Top in vivo essential genes included purA, encoding adenylsuccinate synthetase, and the cps operon required for capsule production. Surprisingly, CRISPRi-seq indicated no fitness-related role for pneumolysin during superinfection. Interestingly, although metK (encoding S-adenosylmethionine synthetase) was essential in vitro, it was dispensable in vivo. This highlights advantages of CRISPRi-seq over transposon-based genetic screens, as all genes, including essential genes, can be tested for pathogenesis potential.
Keywords
CRISPRi-seq, Streptococcus pneumoniae, bacterial pathogenesis, bottleneck, influenza A virus superinfection, pneumonia
Pubmed
Web of science
Create date
30/10/2020 11:21
Last modification date
16/02/2021 6:26
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