Department of Cell, Developmental and Regenerative Medicine, Icahn School of Medicine at Mount Sinai, One Gustave L. Levy Place Box 1020 New York, NY 10029-6574, USA.
Department of Developmental and Molecular Biology, Albert Einstein College of Medicine, Bronx NY 10461, USA.
Development. 2021 Jun 15;148(12). doi: 10.1242/dev.198010. Epub 2021 Jun 18.
Maternally provided gene products regulate the earliest events of embryonic life, including formation of the oocyte that will develop into an egg, and eventually into an embryo. Forward genetic screens have provided invaluable insights into the molecular regulation of embryonic development, including the essential contributions of some genes whose products must be provided to the transcriptionally silent early embryo for normal embryogenesis, called maternal-effect genes. However, other maternal-effect genes are not accessible due to their essential zygotic functions during embryonic development. Identifying these regulators is essential to fill the large gaps in our understanding of the mechanisms and molecular pathways contributing to fertility and to maternally regulated developmental processes. To identify these maternal factors, it is necessary to bypass the earlier requirement for these genes so that their potential later functions can be investigated. Here, we report reverse genetic systems to identify genes with essential roles in zebrafish reproductive and maternal-effect processes. As proof of principle and to assess the efficiency and robustness of mutagenesis, we used these transgenic systems to disrupt two genes with known maternal-effect functions: kif5ba and bucky ball.
母体提供的基因产物调控胚胎生命的最早事件，包括卵母细胞的形成，卵母细胞最终将发育成卵子，并进一步发育成胚胎。正向遗传学筛选为胚胎发育的分子调控提供了宝贵的见解，包括一些基因的重要贡献，这些基因的产物必须提供给转录沉默的早期胚胎，以维持正常的胚胎发生，这些基因被称为母体效应基因。然而，由于其在胚胎发育过程中合子的必需功能，其他母体效应基因是无法获得的。鉴定这些调控因子对于填补我们对有助于生育和母体调控发育过程的机制和分子途径的理解中的巨大空白至关重要。为了鉴定这些母体因子，有必要绕过这些基因早期的需求，以便研究它们潜在的后期功能。在这里，我们报告了用于鉴定在斑马鱼生殖和母体效应过程中具有必需作用的基因的反向遗传学系统。作为原理的证明，并评估诱变的效率和稳健性，我们使用这些转基因系统来破坏两个具有已知母体效应功能的基因：kif5ba 和 bucky ball。
