Summary
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.
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Acknowledgments
We thank Dr. Zain Paroo for critical reading of the manuscript and Feng Jiang for technical assistance. Q. L. is a W. A. “Tex” Moncrief Jr. Scholar in Medical Research and a Damon Runyon Scholar supported by the Damon Runyon Cancer Research Foundation (DRS-43). The work is also supported by a Welch grant (I-1608).
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Ye, X., Liu, Q. (2008). Expression, Purification, and Analysis of Recombinant Drosophila Dicer-1 and Dicer-2 Enzymes. In: Barik, S. (eds) RNAi. Methods in Molecular Biology™, vol 442. Humana Press. https://doi.org/10.1007/978-1-59745-191-8_2
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DOI: https://doi.org/10.1007/978-1-59745-191-8_2
Publisher Name: Humana Press
Print ISBN: 978-1-58829-874-4
Online ISBN: 978-1-59745-191-8
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