A simplified miRNA-based gene silencing method for Drosophila melanogaster

Benjamin Haley; David Hendrix; Vinh Trang; Michael Levine

doi:10.1016/j.ydbio.2008.06.015

A simplified miRNA-based gene silencing method for Drosophila melanogaster

Benjamin Haley, David Hendrix, Vinh Trang, Michael Levine

Research output: Contribution to journal › Article › peer-review

66 Scopus citations

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.

Original language	English (US)
Pages (from-to)	482-490
Number of pages	9
Journal	Developmental biology
Volume	321
Issue number	2
DOIs	https://doi.org/10.1016/j.ydbio.2008.06.015
State	Published - Sep 15 2008
Externally published	Yes

All Science Journal Classification (ASJC) codes

Molecular Biology
Cell Biology
Developmental Biology

Keywords

Decapentaplegic
Drosophila
Method
Nelf
RNAi
Transgenic
miRNA
shRNA
siRNA

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10.1016/j.ydbio.2008.06.015

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title = "A simplified miRNA-based gene silencing method for Drosophila melanogaster",

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.",

keywords = "Decapentaplegic, Drosophila, Method, Nelf, RNAi, Transgenic, miRNA, shRNA, siRNA",

author = "Benjamin Haley and David Hendrix and Vinh Trang and Michael Levine",

note = "Funding Information: We thank Heather Melichar, Frederic Biemar, Matt Ronshaugen, and members of the Levine lab for support and advice. We also thank Johannes Bischof and Konrad Basler for supplying the p attB -UAST vector, Norbert Perrimon for the dpp -dsRNA fly stocks, and Mikkiko Siomi for the gift of AGO1 and AGO2 antibodies. B.H. is supported by an American Cancer Society Postdoctoral Fellowship. This work was funded by a grant from the NIH to M.L. (GM34431). ",

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doi = "10.1016/j.ydbio.2008.06.015",

language = "English (US)",

volume = "321",

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T1 - A simplified miRNA-based gene silencing method for Drosophila melanogaster

AU - Haley, Benjamin

AU - Hendrix, David

AU - Trang, Vinh

AU - Levine, Michael

N1 - Funding Information: We thank Heather Melichar, Frederic Biemar, Matt Ronshaugen, and members of the Levine lab for support and advice. We also thank Johannes Bischof and Konrad Basler for supplying the p attB -UAST vector, Norbert Perrimon for the dpp -dsRNA fly stocks, and Mikkiko Siomi for the gift of AGO1 and AGO2 antibodies. B.H. is supported by an American Cancer Society Postdoctoral Fellowship. This work was funded by a grant from the NIH to M.L. (GM34431).

PY - 2008/9/15

Y1 - 2008/9/15

N2 - 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.

AB - 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.

KW - Decapentaplegic

KW - Drosophila

KW - Method

KW - Nelf

KW - RNAi

KW - Transgenic

KW - miRNA

KW - shRNA

KW - siRNA

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M3 - Article

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SN - 0012-1606

VL - 321

SP - 482

EP - 490

JO - Developmental biology

JF - Developmental biology

IS - 2

ER -

A simplified miRNA-based gene silencing method for Drosophila melanogaster

Abstract

All Science Journal Classification (ASJC) codes

Keywords

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