Skip Navigation

This Article
Right arrow Full Text Freely available
Right arrow Print PDF (343K) Freely available
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Add to My Personal Archive
Right arrow Download to citation manager
Right arrowRequest Permissions
Right arrow Commercial Re-use Guidelines
for Open Access NAR Content
Google Scholar
Right arrow Articles by Piccin, A.
Right arrow Articles by Costa, R.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Piccin, A.
Right arrow Articles by Costa, R.
Related Collections
Right arrow Mutagenesis
Right arrow Cell biology
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us  
What's this?

Nucleic Acids Research, 2001, Vol. 29, No. 12 e55
© 2001 Oxford University Press

Efficient and heritable functional knock-out of an adult phenotype in Drosophila using a GAL4-driven hairpin RNA incorporating a heterologous spacer

Alberto Piccin*, Ahmad Salameh, Clara Benna, Federica Sandrelli, Gabriella Mazzotta, Mauro Zordan, Ezio Rosato1, Charalambos P. Kyriacou2 and Rodolfo Costa

Dipartimento di Biologia, Università di Padova, via Ugo Bassi 58/B, 35131 Padova, Italy, 1Department of Biology and 2Department of Genetics, University of Leicester, Leicester LE1 7RH, UK

We have developed a modified RNA interference (RNAi) method for generating gene knock-outs in Drosophila melanogaster. We used the sequence of the yellow (y) locus to construct an inverted repeat that will form a double-stranded hairpin structure (y-IR) that is under the control of the upstream activating sequence (UAS) of the yeast transcriptional activator GAL4. Hairpins are extremely difficult to manipulate in Escherichia coli, so our method makes use of a heterologous 330 bp spacer encoding sequences from green fluorescent protein to facilitate the cloning steps. When the UAS–y–IR hairpin is expressed under the control of different promoter–GAL4 fusions, a high frequency of y pigment phenocopies is obtained in adults. Consequently this method for producing gene knock-outs has several advantages over previous methods in that it is applicable to any gene within the fly genome, greatly facilitates cloning of the hairpin, can be used if required with GAL4 drivers to avoid lethality or to induce RNAi in a specific developmental stage and/or tissue, is useful for generating knock-outs of adult phenotypes as reported here and, finally, the system can be manipulated to investigate the trans-acting factors that are involved in the RNAi mechanism.

* To whom correspondence should be addressed. Tel: +39 049 827 6228; Fax: +39 049 827 6209; Email: costast4{at}civ.bio.unipd.it


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


This article has been cited by other articles:


Home page
 J. Cell Sci.Home page
A. Jung, M. Hollmann, and M. A. Schafer
The fatty acid elongase NOA is necessary for viability and has a somatic role in Drosophila sperm development
J. Cell Sci., August 15, 2007; 120(16): 2924 - 2934.
[Abstract] [Full Text] [PDF]

Home page
 DevelopmentHome page
H. Furuhashi, M. Nakajima, and S. Hirose
DNA supercoiling factor contributes to dosage compensation in Drosophila
Development, November 15, 2006; 133(22): 4475 - 4483.
[Abstract] [Full Text] [PDF]

Home page
 Nucleic Acids ResHome page
A. E. Brown, L. Bugeon, A. Crisanti, and F. Catteruccia
Stable and heritable gene silencing in the malaria vector Anopheles stephensi
Nucleic Acids Res., August 1, 2003; 31(15): e85 - e85.
[Abstract] [Full Text] [PDF]

Home page
 J. Exp. Biol.Home page
A. E. Brown, A. Crisanti, and F. Catteruccia
Comparative analysis of DNA vectors at mediating RNAi in Anopheles mosquito cells and larvae
J. Exp. Biol., June 1, 2003; 206(11): 1817 - 1823.
[Abstract] [Full Text] [PDF]

Home page
 Proc. Natl. Acad. Sci. USAHome page
S. Dzitoyeva, N. Dimitrijevic, and H. Manev
gamma -Aminobutyric acid B receptor 1 mediates behavior-impairing actions of alcohol in Drosophila: Adult RNA interference and pharmacological evidence
PNAS, April 29, 2003; 100(9): 5485 - 5490.
[Abstract] [Full Text] [PDF]


Disclaimer: Please note that abstracts for content published before 1996 were created through digital scanning and may therefore not exactly replicate the text of the original print issues. All efforts have been made to ensure accuracy, but the Publisher will not be held responsible for any remaining inaccuracies. If you require any further clarification, please contact our Customer Services Department.