The DOI matches its Crossref record; 25 of the 25 references it cites were found in a registry or an index.
In vitro reconstitution of the human RISC-loading complex
Open accessAbstract
Targeted gene silencing by RNAi requires the RNA-induced silencing complex (RISC), whose core component is the protein Argonaute (Ago) bound to a microRNA (miRNA) or an siRNA. In humans, Ago2 is loaded with miRNAs by the action of a specialized assembly called the RISC-loading complex (RLC), comprising the proteins Ago2, Dicer, and TRBP. Here we show that the human RLC assembles spontaneously in vitro from purified components. No cofactors or chaperones are required for the complex to form. The reconstituted RLC, containing one copy of each protein, has the dicing, slicing, guide-strand selection, and Ago2-loading activities observed for the endogenous RLC. Furthermore, once Ago2 is loaded with an miRNA, it tends to dissociate from the rest of the complex. These results lay the groundwork for future structural and functional dissection of RISC loading in humans.
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Cites
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Metadata
- Authors
- MacRae, I. J., Ma, E., Zhou, M., Robinson, C. V., & Doudna, J. A.
- Venue
- Proceedings of the National Academy of Sciences
- Published
- 2008-01-15
- Volume
- 105
- Issue
- 2
- Pages
- 512-517
- ISSN
- 0027-8424 (ISSN-L)
- 1091-6490
- OpenAlex
- W1996830006
- Cited by
- 434
- References
- 25
- FWCI
- 14.89
- Type
- journal article
- Language
- en
- Open access
- green
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- MacRae, I. J., Ma, E., Zhou, M., Robinson, C. V., & Doudna, J. A. (2008). In vitro reconstitution of the human RISC-loading complex. Proceedings of the National Academy of Sciences. https://doi.org/10.1073/pnas.0710869105
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