CRISPRi as a Tool to Repress Multiple Copies of Extracellular Polymeric Substances (EPS)-Related Genes in the Cyanobacterium <i>Synechocystis</i> sp. PCC 6803

The use of the versatile cyanobacterial extracellular polymeric substances (EPS) for biotechnological/biomedical applications implies an extensive knowledge of their biosynthetic pathways to improve/control polymer production yields and characteristics. The multiple copies of EPS-related genes, scat...

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Autores principales:	Marina Santos, Catarina C. Pacheco, Lun Yao, Elton P. Hudson, Paula Tamagnini
Formato:	article
Lenguaje:	EN
Publicado:	MDPI AG 2021
Materias:	CRISPRi cyanobacteria <i>Synechocystis</i> extracellular polymeric substances (EPS) KpsM Science Q
Acceso en línea:	https://doaj.org/article/fe1c7722ca9f44368f281ae8ff61104e
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spelling	oai:doaj.org-article:fe1c7722ca9f44368f281ae8ff61104e2021-11-25T18:11:06ZCRISPRi as a Tool to Repress Multiple Copies of Extracellular Polymeric Substances (EPS)-Related Genes in the Cyanobacterium <i>Synechocystis</i> sp. PCC 680310.3390/life111111982075-1729https://doaj.org/article/fe1c7722ca9f44368f281ae8ff61104e2021-11-01T00:00:00Zhttps://www.mdpi.com/2075-1729/11/11/1198https://doaj.org/toc/2075-1729The use of the versatile cyanobacterial extracellular polymeric substances (EPS) for biotechnological/biomedical applications implies an extensive knowledge of their biosynthetic pathways to improve/control polymer production yields and characteristics. The multiple copies of EPS-related genes, scattered throughout cyanobacterial genomes, adds another layer of complexity, making these studies challenging and time-consuming. Usually, this issue would be tackled by generating deletion mutants, a process that in cyanobacteria is also hindered by the polyploidy. Thus, the use of the CRISPRi multiplex system constitutes an efficient approach to addressing this redundancy. Here, three putative <i>Synechocystis</i> sp. PCC 6803 <i>kpsM</i> homologues (<i>slr0977</i>, <i>slr2107</i>, and <i>sll0574</i>) were repressed using this methodology. The characterization of the 3-sgRNA mutant in terms of fitness/growth and total carbohydrates, released and capsular polysaccharides, and its comparison with previously generated single knockout mutants pointed towards Slr0977 being the key KpsM player in <i>Synechocystis</i> EPS production. This work validates CRISPRi as a powerful tool to unravel cyanobacterial complex EPS biosynthetic pathways expediting this type of studies.Marina SantosCatarina C. PachecoLun YaoElton P. HudsonPaula TamagniniMDPI AGarticleCRISPRicyanobacteria<i>Synechocystis</i>extracellular polymeric substances (EPS)KpsMScienceQENLife, Vol 11, Iss 1198, p 1198 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	CRISPRi cyanobacteria <i>Synechocystis</i> extracellular polymeric substances (EPS) KpsM Science Q
spellingShingle	CRISPRi cyanobacteria <i>Synechocystis</i> extracellular polymeric substances (EPS) KpsM Science Q Marina Santos Catarina C. Pacheco Lun Yao Elton P. Hudson Paula Tamagnini CRISPRi as a Tool to Repress Multiple Copies of Extracellular Polymeric Substances (EPS)-Related Genes in the Cyanobacterium <i>Synechocystis</i> sp. PCC 6803
description	The use of the versatile cyanobacterial extracellular polymeric substances (EPS) for biotechnological/biomedical applications implies an extensive knowledge of their biosynthetic pathways to improve/control polymer production yields and characteristics. The multiple copies of EPS-related genes, scattered throughout cyanobacterial genomes, adds another layer of complexity, making these studies challenging and time-consuming. Usually, this issue would be tackled by generating deletion mutants, a process that in cyanobacteria is also hindered by the polyploidy. Thus, the use of the CRISPRi multiplex system constitutes an efficient approach to addressing this redundancy. Here, three putative <i>Synechocystis</i> sp. PCC 6803 <i>kpsM</i> homologues (<i>slr0977</i>, <i>slr2107</i>, and <i>sll0574</i>) were repressed using this methodology. The characterization of the 3-sgRNA mutant in terms of fitness/growth and total carbohydrates, released and capsular polysaccharides, and its comparison with previously generated single knockout mutants pointed towards Slr0977 being the key KpsM player in <i>Synechocystis</i> EPS production. This work validates CRISPRi as a powerful tool to unravel cyanobacterial complex EPS biosynthetic pathways expediting this type of studies.
format	article
author	Marina Santos Catarina C. Pacheco Lun Yao Elton P. Hudson Paula Tamagnini
author_facet	Marina Santos Catarina C. Pacheco Lun Yao Elton P. Hudson Paula Tamagnini
author_sort	Marina Santos
title	CRISPRi as a Tool to Repress Multiple Copies of Extracellular Polymeric Substances (EPS)-Related Genes in the Cyanobacterium <i>Synechocystis</i> sp. PCC 6803
title_short	CRISPRi as a Tool to Repress Multiple Copies of Extracellular Polymeric Substances (EPS)-Related Genes in the Cyanobacterium <i>Synechocystis</i> sp. PCC 6803
title_full	CRISPRi as a Tool to Repress Multiple Copies of Extracellular Polymeric Substances (EPS)-Related Genes in the Cyanobacterium <i>Synechocystis</i> sp. PCC 6803
title_fullStr	CRISPRi as a Tool to Repress Multiple Copies of Extracellular Polymeric Substances (EPS)-Related Genes in the Cyanobacterium <i>Synechocystis</i> sp. PCC 6803
title_full_unstemmed	CRISPRi as a Tool to Repress Multiple Copies of Extracellular Polymeric Substances (EPS)-Related Genes in the Cyanobacterium <i>Synechocystis</i> sp. PCC 6803
title_sort	crispri as a tool to repress multiple copies of extracellular polymeric substances (eps)-related genes in the cyanobacterium <i>synechocystis</i> sp. pcc 6803
publisher	MDPI AG
publishDate	2021
url	https://doaj.org/article/fe1c7722ca9f44368f281ae8ff61104e
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CRISPRi as a Tool to Repress Multiple Copies of Extracellular Polymeric Substances (EPS)-Related Genes in the Cyanobacterium <i>Synechocystis</i> sp. PCC 6803

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