Please cite as: CSH Protocols; 2008; doi:10.1101/pdb.ip51

This Protocol
Right arrow Full Text
Right arrow Update/discuss this protocolDiscussion icon
Right arrow Alert me when this protocol is cited
Right arrow Alert me when comments are published
Right arrow Alert me if a correction is posted
Services
Right arrow Similar protocols in this database
Right arrow Alert me to new releases of protocols
Right arrow Save to Personal Folders
Right arrow Download to citation manager
Right arrow Printer-friendly versionPrinter-friendly version
Citing Articles
Right arrow Citing Articles via HighWire
Google Scholar
Right arrow Articles by Misquitta, L.
Right arrow Articles by Paterson, B. M.
PubMed
Right arrow Articles by Misquitta, L.
Right arrow Articles by Paterson, B. M.
Related Collections
Right arrow Molecular Biology, general
Right arrow RNA
Right arrow RNA, general
Right arrow Vectors
Right arrow Transgenic Technology, general
Right arrow Drosophila Transgenics
Right arrow Developmental Biology
Right arrow DNA Delivery/Gene Transfer
Right arrow DNA Delivery/Gene Transfer, general
Right arrow RNA Interference (RNAi)/siRNA
Right arrow Cell Biology, general
Right arrow Preparation of Macromolecules and Introduction into Cells
Right arrow Genetics, general
Right arrow Laboratory Organisms, general
Right arrow Drosophila
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us   Add to Digg   Add to Reddit   Add to Technorati  
What's this?
Legend icon

information_panelInformation Panel

Drosophila RNA Interference (RNAi) Using a Gal-4 Inducible Transgene Vector

Leonie Misquitta, Qin Wei, and Bruce M. Paterson1

Laboratory of Biochemistry and Molecular Biology, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892, USA

1Corresponding author (bruce.paterson{at}nih.gov)


INTRODUCTION

RNA interference (RNAi) is a powerful method for determining the role of specific genes during Drosophila embryogenesis. This protocol describes a method for RNAi in vivo using tissue-specific Gal-4 transgenes to induce dsRNA synthesis from an upstream activator sequence (UAS) vector. This vector contains the desired exonic inverted sequences representing the target gene (preferably more than 400 bp) separated by a unique spacer, the first intron of the actin 5C gene. The inverted repeats are stable during cloning in E. coli with this intronic spacer and the intron is spliced out to produce an almost perfect dsRNA target for Dicer cleavage and the production of siRNAs.


Add to CiteULike CiteULike   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us   Add to Digg Digg   Add to Reddit Reddit   Add to Technorati Technorati    What's this?


This article has been cited by other articles:


Home page
 CSHL ProtocolsHome page
L. Misquitta, Q. Wei, and B. M. Paterson
Preparation of Double-Stranded RNA for Drosophila RNA Interference (RNAi)
CSH Protocols, February 1, 2008; 2008(3): pdb.prot4916 - pdb.prot4916.
[Abstract] [Full Text]

Home page
 CSHL ProtocolsHome page
L. Misquitta, Q. Wei, and B. M. Paterson
Collection of Drosophila Embryos for RNA Interference (RNAi)
CSH Protocols, February 1, 2008; 2008(3): pdb.prot4917 - pdb.prot4917.
[Abstract] [Full Text]

Home page
 CSHL ProtocolsHome page
L. Misquitta, Q. Wei, and B. M. Paterson
Injection of dsRNA into Drosophila Embryos for RNA Interference (RNAi)
CSH Protocols, February 1, 2008; 2008(3): pdb.prot4918 - pdb.prot4918.
[Abstract] [Full Text]