Department of Interdisciplinary Science and Engineering in Health Systems, Okayama University, 3-1-1 Tsushimanaka, Okayama, 700-8530, Japan.
Department of Animal Science, Graduate of Environmental and Life Science, Okayama University, Okayama, 700-8530, Japan.
Sci Rep. 2023 Aug 11;13(1):13050. doi: 10.1038/s41598-023-40361-9.
Single-cell-specific delivery of small RNAs, such as short hairpin RNA (shRNA) and small noncoding RNAs, allows us to elucidate the roles of specific upregulation of RNA expression and RNAi-mediated gene suppression in early embryo development. The photoinduced cytosolic dispersion of RNA (PCDR) method that we previously reported can introduce small RNAs into the cytosol of photoirradiated cells and enable RNA delivery into a single-cell in a spatiotemporally specific manner. However, the PCDR method has only been applied to planer cultured cells and not to embryos. This study demonstrated that the PCDR method can be utilized for photo-dependent cytosolic shRNA delivery into a single blastomere and for single blastomere-specific RNA interference in mouse embryos. Our results indicate that PCDR is a promising approach for studying the developmental process of early embryogenesis.
单细胞特异性递送小 RNA（如短发夹 RNA(shRNA)和小非编码 RNA）使我们能够阐明特定上调 RNA 表达和 RNAi 介导的基因抑制在早期胚胎发育中的作用。我们之前报道的光诱导细胞质分散（PCDR）方法可以将小 RNA 导入光照射细胞的细胞质中，并使 RNA 以时空特异性的方式递送至单细胞。然而，PCDR 方法仅适用于平面培养的细胞，而不适用于胚胎。本研究表明，PCDR 方法可用于将光依赖性细胞质 shRNA 递送至单个卵裂球，并在小鼠胚胎中进行单个卵裂球特异性 RNA 干扰。我们的结果表明，PCDR 是研究早期胚胎发生发育过程的一种很有前途的方法。
