Abstract
RNA interference (RNAi) is a form of posttranscriptional gene silencing mediated by microRNA (miRNA) and small interfering RNA (siRNA). In Drosophila melanogaster, the RNase III enzymes Dicer-1 and Dicer-2 generate miRNA and siRNA, respectively. We describe the methods for the expression, purification, and analysis of recombinant Dicer-1 and Dicer-2 enzymes. Our studies demonstrate that Dicer-1 and Dicer-2 display different substrate specificities and ATP requirements.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Animals
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Cell Line
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Drosophila Proteins* / genetics
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Drosophila Proteins* / isolation & purification
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Drosophila Proteins* / metabolism
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Drosophila melanogaster / enzymology*
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Drosophila melanogaster / genetics
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Humans
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RNA Helicases* / genetics
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RNA Helicases* / isolation & purification
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RNA Helicases* / metabolism
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RNA Interference
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Recombinant Proteins* / genetics
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Recombinant Proteins* / isolation & purification
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Recombinant Proteins* / metabolism
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Ribonuclease III
Substances
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Drosophila Proteins
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Recombinant Proteins
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DCR-2 protein, Drosophila
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Dcr-1 protein, Drosophila
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Ribonuclease III
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RNA Helicases