Expression, purification, and analysis of recombinant Drosophila Dicer-1 and Dicer-2 enzymes.
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.
Ye
Xuecheng
X
Department of Biochemistry, University of Texas Southwestern Medical Center, Dallas, TX, USA.
Liu
Qinghua
Q
D000818:Animals; D002460:Cell Line; D029721:Drosophila Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D004331:Drosophila melanogaster / Q000201:enzymology* / Q000235:genetics; D006801:Humans; D020365:RNA Helicases / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D034622:RNA Interference; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D043244:Ribonuclease III
D029721:Drosophila Proteins; D011994:Recombinant Proteins; C478069:DCR-2 protein, Drosophila; C515033:Dcr-1 protein, Drosophila; D043244:Ribonuclease III; D020365:RNA Helicases