Effective protein inhibition in intact mouse oocytes through peptide nanoparticle-mediated antibody transfection.
Effective protein inhibition in intact mouse oocytes through peptide nanoparticle-mediated antibody transfection.
复制标题
通过肽纳米颗粒介导的抗体转染有效抑制完整小鼠卵母细胞的蛋白质
作者：
Li R;Jin Z;Gao L;Liu P;Yang Z;Zhang D
雌性减数分裂是生殖医学的一个基本研究领域，而小鼠卵母细胞体外成熟模型是研究雌性减数分裂最常用的方法。为了研究一种未知蛋白质在雌性减数分裂中的可能作用(S)，传统的方法是向小鼠卵母细胞中微量注射一种特定的抗体。近年来，在体细胞研究中，多肽纳米颗粒介导的抗体转染法因其高效、低毒、稳定性好、血清相容性强等优点而成为一种流行的工具。然而，到目前为止，还没有研究人员尝试在小鼠卵母细胞上使用这项技术，因为卵母细胞膜(卵黄膜)周围的透明带通常被认为或被证明是抗体和蛋白质等大分子的坚硬屏障。因此，我们尝试使用多肽纳米颗粒将抗体引入小鼠卵母细胞。在这里，我们首次展示了利用我们优化的方法，抗体可以有效地输送到小鼠卵母细胞中，并以高特异性抑制其目标蛋白。我们使用小的GTP酶Arl2作为测试对象蛋白，获得了显著的结果。对于小鼠卵母细胞中未知蛋白的初步功能研究，我们建议多肽纳米颗粒介导的抗体转染法是抗体显微注射的一种更好的替代方法。
Female meiosis is a fundamental area of study in reproductive medicine, and the mouse oocyte model of in vitro maturation (IVM) is most widely used to study female meiosis. To investigate the probable role(s) of an unknown protein in female meiosis, the method traditionally used involves microinjecting a specific antibody into mouse oocytes. Recently, in studies on somatic cells, peptide nanoparticle-mediated antibody transfection has become a popular tool because of its high efficiency, low toxicity, good stability, and strong serum compatibility. However, untill now no researchers have tried using this technique on mouse oocytes because the zona pellucida surrounding the oocyte membrane (vitelline membrane) is usually thought or proved to be a tough barrier to macromolecules such as antibodies and proteins. Therefore, we attempted to introduce an antibody into mouse oocytes using a peptide nanoparticle. Here we show for the first time that with our optimized method, an antibody can be effectively delivered into mouse oocytes and inhibit its target protein with high specificity. We obtained significant results using small GTPase Arl2 as a test subject protein. We propose peptide nanoparticle-mediated antibody transfection to be a superior alternative to antibody microinjection for preliminary functional studies of unknown proteins in mouse oocytes.