运用反义寡核苷酸研究非洲爪蛙母源基因的功能

Study the Function of Maternal Genes Using Antisense Oligonucleiotide and Host Transfer Technique

母源基因在动物胚胎早期发育中的功能是发育生物学研究领域的难点之一。在过去二十多年里,反义寡核苷酸介导的反向遗传学技术在多种模式生物中均得到广泛应用,在非洲爪蛙母源基因功能分析中,反义寡核苷酸介导的mRNA降解和受体转移(host transfer)技术的联合使用,确立了以转录因子VegT和Wnt信号分子家族成员Wnt11为代表的母源基因在非洲爪蛙胚胎早期图式建成中的重要功能。目前,非洲爪蛙仍然是研究脊椎动物母源基因功能最为方便的模式系统,因此,该文将在简要叙述VegT和Wnt11调控胚胎三胚层的决定与分化和背方组织者中心诱导过程中的作用的基础上,较为详细地介绍反义脱氧寡核苷酸降解母源mRNA的原理和其与受体转移技术结合使用,分析非洲爪蛙母源基因功能的详细技术流程。

Xenopus laevis has been an important model organism for the study of fundamental questions in molecular biology, cell biology, membrane physiology and developmental biology. In the past two to three decades, Xenopus has been demonstrated as the most amenable vertebrate model system for deciphering the function of ma- ternal genes. By the combinatorial use of antisense oligodeoxynucleotide-mediated mRNA degradation and host transfer techniques, maternal transcriptional factor VegT has been demonstrated to play essential roles in the mesen- doderm induction and maternal Wntl 1 as a critical component of the maternal dorsal determinants. In this review, we will introduce the principles related to the design and use of antisense oligodeoxynucleotide for the depletion of maternal mRNA of interest and the detailed procedures of host transfer technique.