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PMID: 18598689 Published · ppublish English Comparative Study Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't

A simplified miRNA-based gene silencing method for Drosophila melanogaster.

Developmental biology ·Vol. 321 ·No. 2 ·2008-09-15 ·Pages 482-90

Haley B, Hendrix D, Trang V, Levine M

Abstract

MicroRNA-based RNA interference is commonly used to produce loss-of-function phenotypes in mammalian systems, but is used only sparingly in invertebrates such as Caenorhabditis elegans and Drosophila melanogaster. Here, we evaluate this method in transgenic strains of D. melanogaster and cultured S2 cells. High throughput-ready expression vectors were developed that permit rapid cloning of synthetic hairpin RNAs. As proof of concept, this method was used for the efficient silencing of dpp gene activity in the adult wing, and the analysis of the general RNA Polymerase II (Pol II) elongation factor, Nelf-E.

MeSH Terms
Animals Cell Line Drosophila Proteins/genetics,metabolism Drosophila melanogaster/genetics Genetic Techniques Genetic Vectors/genetics MicroRNAs/genetics RNA Interference RNA-Binding Proteins/genetics,metabolism Transcription Factors Wings, Animal/metabolism
Chemicals
Drosophila Proteins MicroRNAs RNA-Binding Proteins Transcription Factors dpp protein, Drosophila negative elongation factor
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Haley Benjamin
Department of Molecular and Cell Biology, Division of Genetics, Genomics, and Development, Center for Integrative Genomics, University of California, Berkeley, Berkeley, California 94720, USA. benhaley@berkeley.edu
Hendrix David
Trang Vinh
Levine Michael
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Article Info
Journal
Developmental biology
Abbr.
Dev Biol
ISSN
1095-564X
DOI
10.1016/j.ydbio.2008.06.015
Email
benhaley@berkeley.edu
Published
2008-09-15
Epub
2008-00-19
Pages
482-90
Language
English
Region
United States
NLM ID
0372762
PMCID
PMC2661819
Subset
IM
Grants
NIGMS NIH HHS · R01 GM034431 · United States
NIGMS NIH HHS · R01 GM034431-24 · United States
NIGMS NIH HHS · GM34431 · United States
External Links
PubMed source DOI: 10.1016/j.ydbio.2008.06.015 PMC Full Text View journal details
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