Mobile-CRISPRi as a powerful tool for modulating Vibrio gene expression.
bioRxiv
; 2024 Jan 18.
Article
em En
| MEDLINE
| ID: mdl-38293084
ABSTRACT
CRISPRi (Clustered Regularly Interspaced Palindromic Repeats interference) is a gene knockdown method that uses a deactivated Cas9 protein (dCas9) that binds a specific gene target locus dictated by an encoded guide RNA (sgRNA) to block transcription. Mobile-CRISPRi is a suite of modular vectors that enable CRISPRi knockdowns in diverse bacteria by integrating IPTG-inducible dcas9 and sgRNA genes into the genome using Tn 7 transposition. Here, we show that the Mobile-CRISPRi system functions robustly and specifically in multiple Vibrio species Vibrio cholerae, Vibrio fischeri, Vibrio vulnificus, Vibrio parahaemolyticus , and Vibrio campbellii . We demonstrate efficacy by targeting both essential and non-essential genes that function to produce defined, measurable phenotypes bioluminescence, quorum sensing, cell division, and growth arrest. We anticipate that Mobile-CRISPRi will be used in Vibrio species to systematically probe gene function and essentiality in various behaviors and native environments.
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MEDLINE
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En
Ano de publicação:
2024
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Article