Efficient Generation of Gene-Modified Pigs HarboringPrecise Orthologous Human Mutation via CRISPRCas

• 作 者: Xiaoyang Zhou Lulu Wang Yinan Du Fei Xie Liang Li Yu Liu Chuanhong Liu Shiqiang Wang Shibing Zhang Xingxu Huang Yong Wang Hong Wei
• 机 构: DepartmentofLaboratoryAnimalScience,CollegeofBasicMedicalSciences,ThirdMilitaryMedicalUniversity,Cho
• 出 处: 第十二届中国实验动物科学年会
• 发表日期: 2016年10期
• 关键词: SOX10 CRISPR/Cas9 homology-directed repair pigs model organism disease model

　　Precise genetic mutation of model animals is highly valuable for functional investigation of human mutations.Clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated 9 (Cas9)-induced homology-directed repair (HDR) is usually used for precise genetic mutation,being limited by the relatively low efficiency compared with that of non-homologous end joining (NHEJ).Although inhibition of NHEJ was shown to enhance HDR-derived mutation,in this work,without inhibition of NHEJ,we first generated gene-modified pigs harboring precise orthologous human mutation (Sox10 c.A325≥T) via CRISPR/Cas9-induced HDR in zygotes using single-strand oligo DNA (ssODN) as template with an efficiency as high as 80%,indicating that pig zygotes exhibited high activities of HDR relative to NHEJ and were highly amendable to genetic mutation via CIRSPR/Cas9-induced HDR.