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通过过量表达秀丽隐杆线虫 SID-1 介导的培养家蚕细胞中的有效 RNA 干扰。

Effective RNA interference in cultured silkworm cells mediated by overexpression of Caenorhabditis elegans SID-1.

机构信息

Laboratory of Silkworm Science, Kyushu University Graduate School of Bioresource and Bioenvironmental Sciences, Hakozaki, Fukuoka, Japan.

出版信息

RNA Biol. 2012 Jan;9(1):40-6. doi: 10.4161/rna.9.1.18084. Epub 2012 Jan 1.

Abstract

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.

摘要

RNA 干扰 (RNAi) 是一种保守的机制,可催化序列特异性基因沉默,并已被用于许多生物体的功能丧失遗传筛选。在这里,我们证明秀丽隐杆线虫 SID-1(CeSID-1)的表达可以在培养的家蚕细胞系 BmN4(BmN4-SID1)中引发有效的基因沉默。将 BmN4-SID1 浸泡在与内源性靶基因对应的 dsRNA 中会导致 mRNA 或蛋白质的量显著减少。少量的 dsRNA 足以在几天内沉默靶基因。CeSID-1 的过表达不会影响细胞活力。我们的结果表明,BmN4-SID1 可在家蚕细胞的许多应用中使用,并将成为基因分析的宝贵资源。

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