The genetic program Of oocytes can be modified in vivo in the zebrafish ovary  

作　　者：Xiaotong Wu Weimin Shen Bingjie Zhang Anming Meng 

机构地区：[1]Laboratory of Molecular Developmental Biology,State Key Laboratory of Membrane Biology,Tsinghua-Peking Center for Life Sciences,School of Life Sciences,Tsinghua University,Beijing 100084,China [2]Center for Stem Cell Biology and Regenerative Medicine,MOE Key Laboratory of Bioinformatics,Tsinghua-Peking Center for Life Sciences,School of Life Sciences,Tsinghua University,Beijing 100084,China

出　　处：《Journal of Molecular Cell Biology》2018年第6期479-493,共15页分子细胞生物学报（英文版）

基　　金：the National Natural Science Foundation of China (31330052 and 31590832).

摘　　要：Oocytes,the irreplaceable gametes for generating a new organism,are matured in the ovary of living female animals.It is unknown whether any genetic manipulations can be applied to immature oocytes inside the living ovaries.As a proof-of-concept, we here demonstrate genetic amendments of zebrafish immature oocytes within the ovary.Oocyte microinjection in situ (OMIS) stimulates tissue repair responses,but some of the microinjected immature oocytes are matured,ovulated and fertilizable.By OMIS-mediated Cas9 approach,ntla and gata5 loci of oocytes arrested at prophase I of meiosis are successfully edited before fertilization.Through OMIS,high efficiency of biaUeUc mutations in single or multiple loci using Cas9/gRNAs allows immediate manifestation of mutant phenotypes in Fo embryos and multiple transgenes can co-express the reporters in Fo embryos with patterns similar to germline transgenic embryos.Furthermore,maternal knockdown of dnmt1 by antisense morphoino via OMIS results in a dramatic decrease of global DNA methylation level at the dome stage and causes embryonic lethalityprior to segmentation period.Therefore,OMIS opens a door to efficiently modify the genome and provides a possibility to re repair genetically abnormal oocytes in situ.

关 键 词：OOCYTES MICROINJECTION gene editing TRANSGENESIS MORPHOLINO ZEBRAFISH 

分 类 号：Q[生物学]