Generation of complement protein C3 deficient pigs by CRISPR/Cas9-mediated gene targeting

Abstract Complement protein C3 is the pivotal component of the complement system. Previous studies have demonstrated that C3 has implications in various human diseases and exerts profound functions under certain conditions. However, the delineation of pathological and physiological roles of C3 has b...

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Autores principales:	Wei Zhang, Guan Wang, Ying Wang, Yong Jin, Lihua Zhao, Qiang Xiong, Lining Zhang, Lisha Mou, Rongfeng Li, Haiyuan Yang, Yifan Dai
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2017
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/3d02ccdbd8c548139931e32b75565b4e
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spelling	oai:doaj.org-article:3d02ccdbd8c548139931e32b75565b4e2021-12-02T16:06:42ZGeneration of complement protein C3 deficient pigs by CRISPR/Cas9-mediated gene targeting10.1038/s41598-017-05400-22045-2322https://doaj.org/article/3d02ccdbd8c548139931e32b75565b4e2017-07-01T00:00:00Zhttps://doi.org/10.1038/s41598-017-05400-2https://doaj.org/toc/2045-2322Abstract Complement protein C3 is the pivotal component of the complement system. Previous studies have demonstrated that C3 has implications in various human diseases and exerts profound functions under certain conditions. However, the delineation of pathological and physiological roles of C3 has been hampered by the insufficiency of suitable animal models. In the present study, we applied the clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated (Cas) system to target the C3 gene in porcine fetal fibroblasts. Our results indicated that CRISPR/Cas9 targeting efficiency was as high as 84.7%, and the biallelic mutation efficiency reached at 45.7%. The biallelic modified colonies were used as donor for somatic cell nuclear transfer (SCNT) technology to generate C3 targeted piglets. A total of 19 C3 knockout (KO) piglets were produced and their plasma C3 protein was undetectable by western blot analysis and ELISA. The hemolytic complement activity and complement-dependent cytotoxicity assay further confirmed that C3 was disrupted in these piglets. These C3 KO pigs could be utilized as a valuable large animal model for the elucidation of the roles of C3.Wei ZhangGuan WangYing WangYong JinLihua ZhaoQiang XiongLining ZhangLisha MouRongfeng LiHaiyuan YangYifan DaiNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 7, Iss 1, Pp 1-9 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Wei Zhang Guan Wang Ying Wang Yong Jin Lihua Zhao Qiang Xiong Lining Zhang Lisha Mou Rongfeng Li Haiyuan Yang Yifan Dai Generation of complement protein C3 deficient pigs by CRISPR/Cas9-mediated gene targeting
description	Abstract Complement protein C3 is the pivotal component of the complement system. Previous studies have demonstrated that C3 has implications in various human diseases and exerts profound functions under certain conditions. However, the delineation of pathological and physiological roles of C3 has been hampered by the insufficiency of suitable animal models. In the present study, we applied the clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated (Cas) system to target the C3 gene in porcine fetal fibroblasts. Our results indicated that CRISPR/Cas9 targeting efficiency was as high as 84.7%, and the biallelic mutation efficiency reached at 45.7%. The biallelic modified colonies were used as donor for somatic cell nuclear transfer (SCNT) technology to generate C3 targeted piglets. A total of 19 C3 knockout (KO) piglets were produced and their plasma C3 protein was undetectable by western blot analysis and ELISA. The hemolytic complement activity and complement-dependent cytotoxicity assay further confirmed that C3 was disrupted in these piglets. These C3 KO pigs could be utilized as a valuable large animal model for the elucidation of the roles of C3.
format	article
author	Wei Zhang Guan Wang Ying Wang Yong Jin Lihua Zhao Qiang Xiong Lining Zhang Lisha Mou Rongfeng Li Haiyuan Yang Yifan Dai
author_facet	Wei Zhang Guan Wang Ying Wang Yong Jin Lihua Zhao Qiang Xiong Lining Zhang Lisha Mou Rongfeng Li Haiyuan Yang Yifan Dai
author_sort	Wei Zhang
title	Generation of complement protein C3 deficient pigs by CRISPR/Cas9-mediated gene targeting
title_short	Generation of complement protein C3 deficient pigs by CRISPR/Cas9-mediated gene targeting
title_full	Generation of complement protein C3 deficient pigs by CRISPR/Cas9-mediated gene targeting
title_fullStr	Generation of complement protein C3 deficient pigs by CRISPR/Cas9-mediated gene targeting
title_full_unstemmed	Generation of complement protein C3 deficient pigs by CRISPR/Cas9-mediated gene targeting
title_sort	generation of complement protein c3 deficient pigs by crispr/cas9-mediated gene targeting
publisher	Nature Portfolio
publishDate	2017
url	https://doaj.org/article/3d02ccdbd8c548139931e32b75565b4e
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Generation of complement protein C3 deficient pigs by CRISPR/Cas9-mediated gene targeting

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