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- W2762296535 abstract "To silence target mRNAs, small RNAs and Argonaute (Ago) proteins need to be assembled into RNA-induced silencing complexes (RISCs). Although the assembly of Drosophila melanogaster RISC was recently reconstituted by Ago2, the Dicer-2/R2D2 heterodimer, and five chaperone proteins, the absence of a reconstitution system for mammalian RISC assembly has posed analytical challenges. Here we describe reconstitution of human RISC assembly using Ago2 and five recombinant chaperone proteins: Hsp90β, Hsc70, Hop, Dnaja2, and p23. Our data show that ATP hydrolysis by both Hsp90β and Hsc70 is required for RISC assembly of small RNA duplexes but not for that of single-stranded RNAs. The reconstitution system lays the groundwork for further studies of small RNA-mediated gene silencing in mammals." @default.
- W2762296535 created "2017-10-20" @default.
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- W2762296535 date "2017-10-02" @default.
- W2762296535 modified "2023-10-12" @default.
- W2762296535 title "In vitro reconstitution of chaperone-mediated human RISC assembly" @default.
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- W2762296535 doi "https://doi.org/10.1261/rna.063891.117" @default.
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