Nakagawa Tatsuya, Emori Chihiro, Ikawa Masahito
Research Institute for Microbial Diseases, Osaka University.
Graduate School of Pharmaceutical Sciences, Osaka University.
Exp Anim. 2025 Jan 11. doi: 10.1538/expanim.24-0165.
In mammals, blastocyst-stage trophectoderm (TE) contacts the maternal body at the time of implantation and forms the placenta after implantation, which supports the development of the fetus. Studying gene function in TE and placenta is important to understand normal implantation and pregnancy processes and their dysfunction. However, genetically modified mice are commonly generated by manipulating pronuclear-stage zygotes, which modify both the genome of the fetus and the placenta. Therefore, we previously developed TE/placenta-specific gene expression technology by transducing blastocysts with lentiviral vectors. However, the zona pellucida (ZP) needed to be removed before transduction. In this study, we examined various adeno-associated viral (AAV) vectors to develop a new TE/placenta-specific gene transduction method. As AAV1 can path through ZP, we succeeded in trophoblast-specific gene expression without ZP removal. Furthermore, TE cells genetically modified by AAV1-Cre contributed uniformly to the placenta. Our new technology contributes to advances in implantation and placenta research and leads to the development of new assisted reproductive technology.
在哺乳动物中,囊胚期滋养外胚层(TE)在着床时与母体组织接触,并在着床后形成胎盘,胎盘支持胎儿发育。研究TE和胎盘中的基因功能对于理解正常的着床和妊娠过程及其功能障碍至关重要。然而,转基因小鼠通常是通过操纵原核期受精卵来产生的,这会同时改变胎儿和胎盘的基因组。因此,我们之前通过用慢病毒载体转导囊胚开发了TE/胎盘特异性基因表达技术。然而,在转导前需要去除透明带(ZP)。在本研究中,我们检测了多种腺相关病毒(AAV)载体,以开发一种新的TE/胎盘特异性基因转导方法。由于AAV1可以穿过ZP,我们成功地在不去除ZP的情况下实现了滋养层特异性基因表达。此外,经AAV1-Cre基因修饰的TE细胞均匀地参与了胎盘的形成。我们的新技术有助于着床和胎盘研究的进展,并推动新的辅助生殖技术的发展。