siRNAs generated by recombinant human Dicer induce specific and significant but target site-independent gene silencing in human cells | Zendy

Hiroaki Kawasaki | Zendy

Open Access

siRNAs generated by recombinant human Dicer induce specific and significant but target site-independent gene silencing in human cells

Author(s) -

Hiroaki Kawasaki

Publication year - 2006

Publication title -

nucleic acids research

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 9.008

H-Index - 537

eISSN - 1362-4954

pISSN - 0305-1048

DOI - 10.1093/nar/gkl306

Subject(s) - biology , dicer , small interfering rna , trans acting sirna , gene silencing , rna interference , microbiology and biotechnology , rna silencing , gene expression , gene , rna induced silencing complex , recombinant dna , rna , genetics

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.

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