Efficient Gene Knockdown in Mouse Oocytes through Peptide Nanoparticle-Mediated SiRNA Transfection.
Efficient Gene Knockdown in Mouse Oocytes through Peptide Nanoparticle-Mediated SiRNA Transfection.
复制标题
通过肽纳米颗粒介导的 siRNA 转染有效敲除小鼠卵母细胞中的基因
DOI：
10.1371/journal.pone.0150462
复制
发表时间：
2016
期刊：
影响因子：
3.7
通讯作者：
Zhang D
中科院分区：
文献类型：
--
作者：
Jin Z;Li R;Zhou C;Shi L;Zhang X;Yang Z;Zhang D
利用小鼠卵母细胞作为研究雌性减数分裂的模型在生殖医学中具有重要意义。在小鼠卵母细胞体外成熟过程中，利用特异性小干扰RNA（siRNA）进行基因敲低通常是研究目的基因功能的第一步。传统上，将siRNA导入小鼠卵母细胞的唯一方法是通过显微注射，这肯定比体细胞中的siRNA转染效率低且费力。近年来，在体细胞研究中，肽纳米颗粒介导的siRNA转染由于其高效、低毒、稳定性好和血清相容性强而比脂质体纳米颗粒介导的方法更受欢迎。然而，由于卵母细胞膜（卵黄膜）周围存在保护性的透明质酸，还没有研究人员尝试将siRNA转染到小鼠卵母细胞中。因此，我们测试了肽纳米颗粒是否可以将siRNA引入小鼠卵母细胞。在本研究中，我们首次表明，我们的优化程序可以有效地敲低靶基因具有高特异性。此外，在敲除测试未知靶基因TRIM75后，我们获得了预期的减数分裂表型。我们建议，肽纳米粒子可能是上级的小鼠卵母细胞中的未知基因的初步功能研究。
The use of mouse oocytes as a model for studying female meiosis is very important in reproductive medicine. Gene knockdown by specific small interfering RNA (siRNA) is usually the first step in the study of the function of a target gene in mouse oocytes during in vitro maturation. Traditionally, the only way to introduce siRNA into mouse oocytes is through microinjection, which is certainly less efficient and strenuous than siRNA transfection in somatic cells. Recently, in research using somatic cells, peptide nanoparticle-mediated siRNA transfection has been gaining popularity over liposome nanoparticle-mediated methods because of its high efficiency, low toxicity, good stability, and strong serum compatibility. However, no researchers have yet tried transfecting siRNA into mouse oocytes because of the existence of the protective zona pellucida surrounding the oocyte membrane (vitelline membrane). We therefore tested whether peptide nanoparticles can introduce siRNA into mouse oocytes. In the present study, we showed for the first time that our optimized program can efficiently knock down a target gene with high specificity. Furthermore, we achieved the expected meiotic phenotypes after we knocked down a test unknown target gene TRIM75. We propose that peptide nanoparticles may be superior for preliminary functional studies of unknown genes in mouse oocytes.