Mobile-CRISPRi protocol optimization for <i>Vibrionaceae</i>-Reference-Cited by-同舟云学术

Mobile-CRISPRi protocol optimization for Vibrionaceae

Published:2024-08-13 Issue:8 Volume:13 Page:
ISSN:2576-098X
Container-title:Microbiology Resource Announcements
language:en
Short-container-title:Microbiol Resour Announc

Author:

Geyman Logan J.¹^ORCID,Tanner Madeline P.¹,Rosario-Meléndez Natalia²³^ORCID,Peters Jason M.³⁴,Mandel Mark J.²³^ORCID,van Kessel Julia C.¹^ORCID

Affiliation:

1. Department of Biology, Indiana University , Bloomington, Indiana, USA

2. Department of Medical Microbiology and Immunology, University of Wisconsin-Madison , Madison, Wisconsin, USA

3. Microbiology Doctoral Training Program, University of Wisconsin-Madison , Madison, Wisconsin, USA

4. Pharmaceutical Sciences Division, School of Pharmacy, University of Wisconsin-Madison , Madison, Wisconsin, USA

Abstract

ABSTRACT Mobile clustered regularly interspaced palindromic repeats interference (Mobile-CRISPRi) is an established method for bacterial gene expression knockdown. The deactivated Cas9 protein and guide RNA are isopropyl β-D-1-thiogalactopyranoside inducible, and all components are integrated into the chromosome via Tn 7 transposition. Here, we optimized methods specific for applying Mobile-CRISPRi in multiple Vibrio species.

Funder

HHS | National Institutes of Health

NSF | National Science Foundation Graduate Research Fellowship Program

Publisher

American Society for Microbiology

Link

https://journals.asm.org/doi/pdf/10.1128/mra.00040-24

Reference6 articles.

1. Diversity in Natural Transformation Frequencies and Regulation across Vibrio Species

2. Enabling genetic analysis of diverse bacteria with Mobile-CRISPRi

3. Mobile-CRISPRi as a powerful tool for modulating Vibrio gene expression

4. A general system to integratelacZ fusions into the chromosomes of gram-negative eubacteria: regulation of thePm promoter of theTOL plasmid studied with all controlling elements in monocopy

5. RP4-based plasmids for conjugation between Escherichia coli and members of the vibrionaceae