Modification of alternative splicing in bovine somatic cell nuclear transfer embryos using engineered CRISPR-Cas13d

作     者：Rui Cheng Xiaoman Zheng Yingmei Wang Xing Ma Xin Liu Wenjun Xu Mengyun Wang Yuanpeng Gao Xupeng Xing Chuan Zhou Hongzheng Sun Zekun Guo Fusheng Quan Jun Liu Song Hua Yongsheng Wang Yong Zhang Xu Liu 

作者机构：College of Veterinary MedicineNorthwest A&F UniversityKey Laboratory of Animal Biotechnology of the Ministry of AgricultureYangling 712100China 

出 版 物：《Science China(Life Sciences)》 (中国科学（生命科学英文版）)

年 卷 期：2022年第65卷第11期

页      面：2257-2268页

核心收录：

学科分类：0710[理学-生物学] 07[理学] 0905[农学-畜牧学] 08[工学] 09[农学] 071007[理学-遗传学] 0901[农学-作物学] 0836[工学-生物工程] 090102[农学-作物遗传育种] 

基　　金：the National Natural Science Foundation of China(31802153,32072805 and 31872539) Special Funds for Talents in Northwest A&F University(Z111021512 and Z109021702) Innovation Project of Science and Technology in Shaanxi Province(2018JQ3035) National Major Project for Production of Transgenic Breeding(2016ZX08007-003) 

主　　题：animal cloning alternative splicing CRISPR-Cas13d bovine 

摘      要：Animal cloning can be achieved by somatic cell nuclear transfer(SCNT),but the resulting live birth rate is relatively *** previously improved the efficiency of bovine SCNT by exogenous melatonin treatment or by overexpression of lysine-specific demethylase 4D(KDM4D)and 4E(KDM4E).In this study,we revealed abundant alternative splicing(AS)transitions during fertilization and embryonic genome activation,and demonstrated abnormal AS in bovine SCNT embryos compared with in vitro fertilized *** used the CRISPR-Cas13d RNA-targeting system to target cis-elements of ABI2 and ZNF106 pre-m RNA to modify AS,thus reducing the ratio of abnormal-isoform SCNT embryos by nearly 50%and achieving a high survival rate(11%–19%).These results indicate that this system may provide an efficient method for bovine cloning,while also paving the way for further improvements in the efficiency of SCNT.