Reconstitution of human RNA interference in budding yeast.
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.
Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, MA, USA. ksuk@knu.ac.kr
Kyoungho
K
Suk
Jihye
J
Choi
Yo
Y
Suzuki
Sedide B
SB
Ozturk
Joseph C
JC
Mellor
Koon Ho
KH
Wong
Joanna L
JL
MacKay
Richard I
RI
Gregory
Frederick P
FP
Roth
D060565:Argonaute Proteins; D015852:Eukaryotic Initiation Factor-2 / Q000235:genetics / Q000378:metabolism; D006801:Humans; D008957:Models, Genetic; D034622:RNA Interference; D016601:RNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D043244:Ribonuclease III / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000378:metabolism; D000099253:Trans-Activation Responsive RNA-Binding Protein
D060565:Argonaute Proteins; D015852:Eukaryotic Initiation Factor-2; D016601:RNA-Binding Proteins; D043244:Ribonuclease III; D000099253:Trans-Activation Responsive RNA-Binding Protein; C466422:AGO2 protein, human
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