Fast cloning inverted repeats for RNA interference

Sujin Bao, Ross Cagan

Research output: Contribution to journalArticlepeer-review

20 Scopus citations

Abstract

Double-stranded RNA (dsRNA) can induce post-transcriptional gene silencing in a wide variety of organisms. Commonly, inverted repeats are used to produce dsRNA to silence genes of interest. However, cloning inverted repeats still remains a rate-limiting step for widely applying this technique. Here we describe a pGEM-T-based vector, pGEM-WIZ, designed to produce inverted repeats for any Drosophila gene. pGEM-WIZ has a high efficiency in assembling inverted repeats and the repeats in this vector are stable in regular Escherichia coli strains. Furthermore, we have developed a method for rapid selection of clones with an inverted repeat based on size and relative copy number of the vector with or without an insert. This method further eases the cloning process. The inverted repeat cassette assembled in pGEM-WIZ can be easily transferred to commonly available expression vectors suitable for stably expressing inverted repeats in vitro and in vivo. Published by Cold Spring Harbor Laboratory Press.

Original languageEnglish
Pages (from-to)2020-2024
Number of pages5
JournalRNA
Volume12
Issue number11
DOIs
StatePublished - 2006
Externally publishedYes

Keywords

  • Cloning
  • Drosophila
  • Inverted repeat
  • RNA interference

Fingerprint

Dive into the research topics of 'Fast cloning inverted repeats for RNA interference'. Together they form a unique fingerprint.

Cite this