Title

In vitro analysis of RNA interference in Drosophila melanogaster

GSBS Program

Biochemistry & Molecular Pharmacology

UMMS Affiliation

Department of Biochemistry and Molecular Pharmacology

Date

6-28-2003

Document Type

Article

Medical Subject Headings

Animals; Drosophila; Embryo, Nonmammalian; *Genetic Techniques; *RNA Interference

Disciplines

Life Sciences | Medicine and Health Sciences

Abstract

Double-stranded RNA (dsRNA) triggers the destruction of mRNA sharing sequence with the dsRNA, a phenomenon termed RNA interference (RNAi). The dsRNA is converted by endonucleolytic cleavage into 21- to 23-nt small interfering RNAs (siRNAs), which direct a multiprotein complex, the RNA-induced silencing complex to cleave RNA complementary to the siRNA. RNAi can be recapitulated in vitro in lysates of syncytial blastoderm Drosophila embryos. These lysates reproduce all of the known steps in the RNAi pathway in flies and mammals. Here we explain how to prepare and use Drosophila embryo lysates to dissect the mechanism of RNAi.

Rights and Permissions

Citation: Methods. 2003 Aug;30(4):330-6.

Related Resources

Link to article in PubMed

PubMed ID

12828947