摘要: Pig shows multiple superior characteristics in anatomy, physiology, and genome that have made this species to be more suitable models for human diseases, especially for neurodegenerative diseases, because they have similar cerebral convolutions compared with human neocortex. Recently, CRISPR/Cas9 system shows enormous potential for engineering the pig genome. In this study, we expect to generate human Parkinson's disease pig model using CRISPR/Cas9 system by simultaneously targeting three distinct genomic loci, parkin/DJ-1/PINK1, in Bama miniature pigs. By co-injection of Cas9 mRNA and multiplexing single guide RNAs (sgRNAs) targeting parkin, DJ-1, and PINK1 genes, respectively, into in vivo derived pronuclear embryos, we simultaneously targeted three distinct genomic loci. The gene modified piglets remain healthy and display normal behavior at the age of 10 months. In addition, despite the high number of sgRNAs were employed in the present study, our trio-based whole-genome sequencing analysis suggested that the incidence of off-target events is low. Our results demonstrate that the simplicity, efficiency, and power of the CRISPR/Cas9 system to allow for the modification of multiple genes in pigs and yield results of high medical value.
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期刊:
SCIENTIFIC REPORTS
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分类:
生物学
>>
生物物理学
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引用:
ChinaXiv:201605.00749
(或此版本
ChinaXiv:201605.00749V1)
DOI:10.12074/201605.00749V1
CSTR:32003.36.ChinaXiv.201605.00749.V1
- 推荐引用方式:
Wang, Xianlong,Cao, Chunwei,Huang, Jiaojiao,Yao, Jing,Hai, Tang,Zheng, Qiantao,Wang, Xiao,Zhang, Hongyong,Qin, Guosong,Zhou, Qi,Wang, Hongmei,Zhao, Jianguo,Zheng, Qiantao,Wang, Xiao,Zhang, Hongyong,Zhou, Qi,Wang, Hongmei,Zhao, Jianguo,Cheng, Jinbo,Yuan, Zengqiang,Wang, Yanfang,.One-step generation of triple gene-targeted pigs using CRISPR/Cas9 system.中国科学院科技论文预发布平台.[ChinaXiv:201605.00749V1]
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