An easy-to-use CRISPRi plasmid tool for inducible knockdown in E. coli

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 mini...

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Detalles Bibliográficos
Autor principal: Bradley, Robert W.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2021
Materias:
Short Communication
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8524100/
https://www.ncbi.nlm.nih.gov/pubmed/34703773
http://dx.doi.org/10.1016/j.btre.2021.e00680
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author Bradley, Robert W.
author_facet Bradley, Robert W.
author_sort Bradley, Robert W.
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description 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.
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spelling pubmed-85241002021-10-25 An easy-to-use CRISPRi plasmid tool for inducible knockdown in E. coli Bradley, Robert W. Biotechnol Rep (Amst) Short Communication 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. Elsevier 2021-10-09 /pmc/articles/PMC8524100/ /pubmed/34703773 http://dx.doi.org/10.1016/j.btre.2021.e00680 Text en © 2021 The Author. Published by Elsevier B.V. https://creativecommons.org/licenses/by/4.0/This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Short Communication
Bradley, Robert W.
An easy-to-use CRISPRi plasmid tool for inducible knockdown in E. coli
title An easy-to-use CRISPRi plasmid tool for inducible knockdown in E. coli
title_full An easy-to-use CRISPRi plasmid tool for inducible knockdown in E. coli
title_fullStr An easy-to-use CRISPRi plasmid tool for inducible knockdown in E. coli
title_full_unstemmed An easy-to-use CRISPRi plasmid tool for inducible knockdown in E. coli
title_short An easy-to-use CRISPRi plasmid tool for inducible knockdown in E. coli
title_sort easy-to-use crispri plasmid tool for inducible knockdown in e. coli
topic Short Communication
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8524100/
https://www.ncbi.nlm.nih.gov/pubmed/34703773
http://dx.doi.org/10.1016/j.btre.2021.e00680
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