CRISPR/Cas12a‐mediated genome engineering in the photosynthetic bacterium Rhodobacter capsulatus

Summary Purple non‐sulfur photosynthetic bacteria (PNSB) such as Rhodobacter capsulatus serve as a versatile platform for fundamental studies and various biotechnological applications. In this study, we sought to develop the class II RNA‐guided CRISPR/Cas12a system from Francisella novicida for geno...

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Autores principales:	Yang Zhang, Jifeng Yuan
Formato:	article
Lenguaje:	EN
Publicado:	Wiley 2021
Materias:	Biotechnology TP248.13-248.65
Acceso en línea:	https://doaj.org/article/bf4cae380400472580b5d016e8379a68
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spelling	oai:doaj.org-article:bf4cae380400472580b5d016e8379a682021-11-18T15:39:53ZCRISPR/Cas12a‐mediated genome engineering in the photosynthetic bacterium Rhodobacter capsulatus1751-791510.1111/1751-7915.13805https://doaj.org/article/bf4cae380400472580b5d016e8379a682021-11-01T00:00:00Zhttps://doi.org/10.1111/1751-7915.13805https://doaj.org/toc/1751-7915Summary Purple non‐sulfur photosynthetic bacteria (PNSB) such as Rhodobacter capsulatus serve as a versatile platform for fundamental studies and various biotechnological applications. In this study, we sought to develop the class II RNA‐guided CRISPR/Cas12a system from Francisella novicida for genome editing and transcriptional regulation in R. capsulatus. Template‐free disruption method mediated by CRISPR/Cas12a reached ˜ 90% editing efficiency when targeting ccoO or nifH gene. When both genes were simultaneously edited, the multiplex editing efficiency reached > 63%. In addition, CRISPR interference (CRISPRi) using deactivated Cas12a was also evaluated using reporter genes egfp and lacZ, and the transcriptional repression efficiency reached ˜ 80%. In summary, our work represents the first report to develop CRISPR/Cas12a‐mediated genome editing and transcriptional regulation in R. capsulatus, which would greatly accelerate PNSB‐related researches.Yang ZhangJifeng YuanWileyarticleBiotechnologyTP248.13-248.65ENMicrobial Biotechnology, Vol 14, Iss 6, Pp 2700-2710 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Biotechnology TP248.13-248.65
spellingShingle	Biotechnology TP248.13-248.65 Yang Zhang Jifeng Yuan CRISPR/Cas12a‐mediated genome engineering in the photosynthetic bacterium Rhodobacter capsulatus
description	Summary Purple non‐sulfur photosynthetic bacteria (PNSB) such as Rhodobacter capsulatus serve as a versatile platform for fundamental studies and various biotechnological applications. In this study, we sought to develop the class II RNA‐guided CRISPR/Cas12a system from Francisella novicida for genome editing and transcriptional regulation in R. capsulatus. Template‐free disruption method mediated by CRISPR/Cas12a reached ˜ 90% editing efficiency when targeting ccoO or nifH gene. When both genes were simultaneously edited, the multiplex editing efficiency reached > 63%. In addition, CRISPR interference (CRISPRi) using deactivated Cas12a was also evaluated using reporter genes egfp and lacZ, and the transcriptional repression efficiency reached ˜ 80%. In summary, our work represents the first report to develop CRISPR/Cas12a‐mediated genome editing and transcriptional regulation in R. capsulatus, which would greatly accelerate PNSB‐related researches.
format	article
author	Yang Zhang Jifeng Yuan
author_facet	Yang Zhang Jifeng Yuan
author_sort	Yang Zhang
title	CRISPR/Cas12a‐mediated genome engineering in the photosynthetic bacterium Rhodobacter capsulatus
title_short	CRISPR/Cas12a‐mediated genome engineering in the photosynthetic bacterium Rhodobacter capsulatus
title_full	CRISPR/Cas12a‐mediated genome engineering in the photosynthetic bacterium Rhodobacter capsulatus
title_fullStr	CRISPR/Cas12a‐mediated genome engineering in the photosynthetic bacterium Rhodobacter capsulatus
title_full_unstemmed	CRISPR/Cas12a‐mediated genome engineering in the photosynthetic bacterium Rhodobacter capsulatus
title_sort	crispr/cas12a‐mediated genome engineering in the photosynthetic bacterium rhodobacter capsulatus
publisher	Wiley
publishDate	2021
url	https://doaj.org/article/bf4cae380400472580b5d016e8379a68
work_keys_str_mv	AT yangzhang crisprcas12amediatedgenomeengineeringinthephotosyntheticbacteriumrhodobactercapsulatus AT jifengyuan crisprcas12amediatedgenomeengineeringinthephotosyntheticbacteriumrhodobactercapsulatus
_version_	1718420809259679744

CRISPR/Cas12a‐mediated genome engineering in the photosynthetic bacterium Rhodobacter capsulatus

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