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Modification of alternative splicing in bovine somatic cell nuclear transfer embryos using engineered CRISPR-Cas13d 被引量:2
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作者 Rui Cheng Xiaoman Zheng +15 位作者 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 《Science China(Life Sciences)》 SCIE CAS CSCD 2022年第11期2257-2268,共12页
Animal cloning can be achieved by somatic cell nuclear transfer(SCNT),but the resulting live birth rate is relatively low.We previously improved the efficiency of bovine SCNT by exogenous melatonin treatment or by ove... Animal cloning can be achieved by somatic cell nuclear transfer(SCNT),but the resulting live birth rate is relatively low.We 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 embryos.We 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. 展开更多
关键词 animal cloning alternative splicing CRISPR-Cas13d BOVINE
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