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Reconstitution of human RNA interference in budding yeast.

ABSTRACT: Although RNA-mediated interference (RNAi) is a widely conserved process among eukaryotes, including many fungi, it is absent from the budding yeast Saccharomyces cerevisiae. Three human proteins, Ago2, Dicer and TRBP, are sufficient for reconstituting the RISC complex in vitro. To examine whether the introduction of human RNAi genes can reconstitute RNAi in S. cerevisiae, genes encoding these three human proteins were introduced into S. cerevisiae. We observed both siRNA and siRNA- and RISC-dependent silencing of the target gene GFP. Thus, human Ago2, Dicer and TRBP can functionally reconstitute human RNAi in S. cerevisiae, in vivo, enabling the study and use of the human RNAi pathway in a facile genetic model organism.

SUBMITTER: Suk K

PROVIDER: S-EPMC3074155 | biostudies-literature | 2011 Apr

REPOSITORIES: biostudies-literature

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Reconstitution of human RNA interference in budding yeast.

Suk Kyoungho K Choi Jihye J Suzuki Yo Y Ozturk Sedide B SB Mellor Joseph C JC Wong Koon Ho KH MacKay Joanna L JL Gregory Richard I RI Roth Frederick P FP

Nucleic acids research 20110119 7

Although RNA-mediated interference (RNAi) is a widely conserved process among eukaryotes, including many fungi, it is absent from the budding yeast Saccharomyces cerevisiae. Three human proteins, Ago2, Dicer and TRBP, are sufficient for reconstituting the RISC complex in vitro. To examine whether the introduction of human RNAi genes can reconstitute RNAi in S. cerevisiae, genes encoding these three human proteins were introduced into S. cerevisiae. We observed both siRNA and siRNA- and RISC-depe ...[more]

PMID: 21252293

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