An easy-to-use CRISPRi plasmid tool for inducible knockdown in E. coli
File(s)
Author(s)
Bradley, Robert
Type
Journal Article
Abstract
CRISPR-interference (CRISPRi) is a powerful genetic tool with wide application in bacteria. In this work a single plasmid system is presented which expresses deactivated Cas9 and a short guide RNA sequence from separate inducible promoters, enabling up to 40-fold repression of target genes with minimal leaky repression in the uninduced state. The plasmid is designed for rapid spacer insertion with easy screening for correct clones, and a simple one-day protocol with >99.9% assembly efficiency is described. This plasmid is made available to the research community with a view to facilitating wider use of CRISPRi in bacterial systems.
Date Issued
2021-12
Date Acceptance
2021-09-29
Citation
Biotechnology Reports, 2021, 32, pp.1-4
ISSN
2215-017X
Publisher
Elsevier
Start Page
1
End Page
4
Journal / Book Title
Biotechnology Reports
Volume
32
Copyright Statement
© 2021 The Author. Published by Elsevier B.V. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
License URL
Sponsor
Biotechnology and Biological Sciences Research Council
Identifier
https://www.sciencedirect.com/science/article/pii/S2215017X21000965?via%3Dihub
Grant Number
BB/R009171/1
Subjects
CRISPRi
Inducible repression
One-pot assembly
Publication Status
Published
Date Publish Online
2021-10-09