Effective RNA interference in cultured silkworm cells mediated by overexpression of Caenorhabditis elegans SID-1
Effective RNA interference in cultured silkworm cells mediated by overexpression of Caenorhabditis elegans SID-1
复制标题
DOI：
10.4161/rna.9.1.18084
复制
发表时间：
2012-01-01
期刊：
影响因子：
4.1
通讯作者：
Kusakabe, Takahiro
中科院分区：
文献类型：
--
作者：
Mon, Hiroaki;Kobayashi, Isao;Kusakabe, Takahiro
RNA干扰（RNAi）是一种保守的催化序列特异性基因沉默的机制，并已被用于许多生物体中的功能丧失遗传筛选。在这里，我们证明了秀丽隐杆线虫SID-1（CeSID-1）的表达可以在培养的家蚕细胞系BmN 4（BmN 4-SID 1）中触发有效的基因沉默。将BmN 4-SID 1浸泡在对应于内源性靶基因的dsRNA中诱导mRNA或蛋白质的量显著降低。少量的dsRNA就足以在几天内使靶基因沉默。CeSID-1的过表达不影响细胞活力。我们的研究结果表明，BmN 4-SID 1可以在家蚕细胞中的许多应用，并将成为一个有价值的资源，基因分析。
RNA interference (RNAi) is a conserved mechanism that catalyzes sequence-specific gene silencing, and has been used for loss-of-function genetic screens in many organisms. Here, we demonstrated that the expression of Caenorhabditis elegans SID-1 (CeSID-1) could trigger effective gene silencing in the cultured silkworm cell line, BmN4 (BmN4-SID1). Soaking the BmN4-SID1 in dsRNA corresponding to endogenous target genes induced a significant decrease of the amount of mRNA or protein. A small amount of dsRNA was enough to silence the target gene in a few days. Overexpression of CeSID-1 did not affect the cell viability. Our results suggest that BmN4-SID1 can be used in many applications in silkworm cells and will become a valuable resource for gene analysis.