siRNAs generated by recombinant human Dicer induce specific and significant but target site-independent gene silencing in human cells.
siRNAs generated by recombinant human Dicer induce specific and significant but target site-independent gene silencing in human cells.
复制标题
重组人 Dicer 产生的 siRNA 在人类细胞中诱导特异性且显着但与靶位点无关的基因沉默。
作者：
H. Kawasaki;E. Suyama;Mayu Iyo;K. Taira
RNA干扰已成为动物和植物基因表达沉默的有力工具。最近有报道称，21nt合成的小干扰rna （sirna）特异性抑制了几种哺乳动物细胞中内源基因的表达。然而，sirna的功效依赖于目标mRNA中特定靶位点的存在，并且仍然很难预测最佳或最有效的靶位点。在这项研究中，我们证明了重组人Dicer （re-hDicer）在体外产生的sirna不仅能显著抑制外源性嘌呤霉素耐药基因的表达，还能抑制内源性H-ras、c-jun和c-fos的表达。在我们的系统中，目标位点的选择在sirna的设计中是不必要的。然而，重要的是要避免同源序列的目标mRNA在一个给定的蛋白质家族。我们的siRNA切割系统应该是一个强大的工具，在哺乳动物细胞基因失活。
RNA interference has emerged as a powerful tool for the silencing of gene expression in animals and plants. It was reported recently that 21 nt synthetic small interfering RNAs (siRNAs) specifically suppressed the expression of endogenous genes in several lines of mammalian cells. However, the efficacy of siRNAs is dependent on the presence of a specific target site within the target mRNA and it remains very difficult to predict the best or most effective target site. In this study, we demonstrate that siRNAs that have been generated in vitro by recombinant human Dicer (re-hDicer) significantly suppress not only the exogenous expression of a puromycin-resistance gene but also the endogenous expression of H-ras, c-jun and c-fos. In our system, selection of a target site is not necessary in the design of siRNAs. However, it is important to avoid homologous sequences within a target mRNA in a given protein family. Our diced siRNA system should be a powerful tool for the inactivation of genes in mammalian cells.
DOI：
10.1073/pnas.95.25.14687
发表时间：
1998-12-08
影响因子：
11.1
作者：
Ngô, H;Tschudi, C;Ullu, E
通讯作者：
Ullu, E