Generation of tryptophan hydroxylase 2 gene knockout pigs by CRISPR/Cas9-mediated gene targeting
Ze Li
Hai-Yuan Yang
Ying Wang
Man-Ling Zhang
Xiao-Rui Liu
Qiang Xiong
Li-Ning Zhang
Yong Jin
Li-Sha Mou
Yan Liu
Rong-Feng Li
Yi Rao
Yi-Fan Dai
摘要：Unbalanced brain serotonin (5-HT) levels have implications in various behavioral abnormalities and neuropsychiatric disorders.The biosynthesis of neuronal 5-HT is regulated by the rate-limiting enzyme,tryptophan hydroxylase-2 (TPH2).In the present study,the clustered regularly interspaced short palindromic repeat (CRISPR)/ CRISPR-associated (Cas) system was used to target the Tph2 gene in Bama mini pig fetal fibroblasts.It was found that CRISPR/Cas9 targeting efficiency could be as high as 61.5％,and the biallelic mutation efficiency reached at 38.5％.The biallelic modified colonies were used as donors for somatic cell nuclear transfer (SCNT) and 10 Tph2 targeted piglets were successfully generated.These Tph2 KO piglets were viable and appeared normal at the birth.However,their central 5-HT levels were dramatically reduced,and their survival and growth rates were impaired before weaning.These Tph2 KO pigs are valuable large-animal models for studies of 5-HT deficiency induced behavior abnomality.
机标关键词：gene knockoutnuclear transfergrowth rates
分类号：R284(中药学)
资助基金：a grant from the National Natural Science Foundation of China(81570402)a grant from the Jiangsu Key Laboratory of Xenotransplantation(BM2012116)grants from the Sanming Project of Medicine in Shenzhen,the Fund for High Level Medical Discipline Construction of Shenzhen(2016031638)the Shenzhen Foundation of Science and Technology(JCYJ20160229204849-975 and GCZX2015043017281705)grant from the National Basic Research Program of China(2015CB55-9200)
论文发表日期：2017-01-01
在线出版日期：2025-08-15（本平台首次上网日期，不代表文献的发表时间）
页数：8( 445-452 )
英文信息