Laboratory of Silkworm Science, Kyushu University Graduate School of Bioresource and Bioenvironmental Sciences, 6-10-1 Hakozaki Higashi-ku, Fukuoka, 812-8581, Japan.
Mol Biotechnol. 2014 Mar;56(3):193-8. doi: 10.1007/s12033-013-9694-0.
The systemic RNA interference defective-1 (SID-1) can transport double-stranded RNA (dsRNA) into cytosol across the cytoplasmic membrane. We report here that ectopic expression of Caenorhabditis elegans SID-1 allows BmN4 cells to import extracellular plasmid dsDNA into cells via the direct soaking method. Interestingly, BmN4-SID1 cells incorporate dsRNA and plasmid DNA simultaneously. Furthermore, the ectopic SID-1 allows us to establish a stably transformed cell line by the simple soaking method. Our results provide an alternative method for silkworm gene function analysis with low cost and low cell toxicity.
系统性 RNA 干扰缺陷-1 (SID-1) 可以将双链 RNA (dsRNA) 通过细胞质膜运送到细胞质中。我们在这里报告，线虫 SID-1 的异位表达允许 BmN4 细胞通过直接浸泡法将细胞外质粒 dsDNA 导入细胞。有趣的是，BmN4-SID1 细胞同时摄取 dsRNA 和质粒 DNA。此外，异位 SID-1 允许我们通过简单的浸泡法建立稳定转化的细胞系。我们的结果提供了一种低成本、低细胞毒性的替代方法，用于分析家蚕基因功能。
