Header

UZH-Logo

Maintenance Infos

A versatile platform for creating a comprehensive UAS-ORFeome library in Drosophila


Bischof, Johannes; Björklund, Mikael; Furger, Edy; Schertel, Claus; Taipale, Jussi; Basler, Konrad (2013). A versatile platform for creating a comprehensive UAS-ORFeome library in Drosophila. Development, 140(11):2434-2442.

Abstract

Overexpression screens are used to explore gene functions in Drosophila, but this strategy suffers from the lack of comprehensive and systematic fly strain collections and efficient methods for generating such collections. Here, we present a strategy that could be used efficiently to generate large numbers of transgenic Drosophila strains, and a collection of 1149 UAS-ORF fly lines that were created with the site-specific ΦC31 integrase method. For this collection, we used a set of 655 genes that were cloned as two variants, either as an open reading frame (ORF) with a native stop codon or with a C-terminal 3xHA tag. To streamline the procedure for transgenic fly generation, we demonstrate the utility of injecting pools of plasmids into embryos, each plasmid containing a randomised sequence (barcode) that serves as a unique identifier for plasmids and, subsequently, fly strains. We also developed a swapping technique that facilitates the rapid exchange of promoters and epitope tags in vivo, expanding the versatility of the ORF collection. The work described here serves as the basis of a systematic library of Gal4/UAS-regulated transgenes.

Abstract

Overexpression screens are used to explore gene functions in Drosophila, but this strategy suffers from the lack of comprehensive and systematic fly strain collections and efficient methods for generating such collections. Here, we present a strategy that could be used efficiently to generate large numbers of transgenic Drosophila strains, and a collection of 1149 UAS-ORF fly lines that were created with the site-specific ΦC31 integrase method. For this collection, we used a set of 655 genes that were cloned as two variants, either as an open reading frame (ORF) with a native stop codon or with a C-terminal 3xHA tag. To streamline the procedure for transgenic fly generation, we demonstrate the utility of injecting pools of plasmids into embryos, each plasmid containing a randomised sequence (barcode) that serves as a unique identifier for plasmids and, subsequently, fly strains. We also developed a swapping technique that facilitates the rapid exchange of promoters and epitope tags in vivo, expanding the versatility of the ORF collection. The work described here serves as the basis of a systematic library of Gal4/UAS-regulated transgenes.

Statistics

Citations

Dimensions.ai Metrics
77 citations in Web of Science®
70 citations in Scopus®
77 citations in Microsoft Academic
Google Scholar™

Altmetrics

Downloads

167 downloads since deposited on 20 Jun 2013
5 downloads since 12 months
Detailed statistics

Additional indexing

Item Type:Journal Article, refereed, original work
Communities & Collections:07 Faculty of Science > Institute of Molecular Life Sciences
Dewey Decimal Classification:570 Life sciences; biology
Language:English
Date:1 May 2013
Deposited On:20 Jun 2013 12:02
Last Modified:23 Sep 2018 05:34
Publisher:Company of Biologists
ISSN:0950-1991
OA Status:Hybrid
Publisher DOI:https://doi.org/10.1242/dev.088757
PubMed ID:23637332

Download

Download PDF  'A versatile platform for creating a comprehensive UAS-ORFeome library in Drosophila'.
Preview
Content: Published Version
Filetype: PDF
Size: 1MB
View at publisher