Expression, Purification, and Analysis of Recombinant Drosophila Dicer-1 and Dicer-2 Enzymes
- 1 January 2008
- book chapter
- Published by Springer Nature
- Vol. 442, 11-27
- https://doi.org/10.1007/978-1-59745-191-8_2
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.Keywords
This publication has 19 references indexed in Scilit:
- Oncomirs — microRNAs with a role in cancerNature Reviews Cancer, 2006
- Micro RNAs in Animal DevelopmentCell, 2006
- Structural Basis for Double-Stranded RNA Processing by DicerScience, 2006
- Silence from within: Endogenous siRNAs and miRNAsCell, 2005
- Dicer-1 and R3D1-L catalyze microRNA maturation in DrosophilaGenes & Development, 2005
- Post-transcriptional gene silencing by siRNAs and miRNAsPublished by Elsevier ,2005
- Processing of Pre-microRNAs by the Dicer-1–Loquacious Complex in Drosophila CellsPLoS Biology, 2005
- RNA interferenceNature, 2002
- Dicer functions in RNA interference and in synthesis of small RNA involved in developmental timing in C. elegansGenes & Development, 2001
- Role for a bidentate ribonuclease in the initiation step of RNA interferenceNature, 2001