A versatile in vivo system for directed dissection of gene expression patterns

Autoren

Daryl M Gohl
Marion A Silies
Xiaojing J Gao
Sheetal Bhalerao
Francisco J Luongo
Chun-Chieh Lin
Christopher J Potter
Thomas R Clandinin

DOI

10.1038/nmeth.1561

eISSN

1548-7105

ISSN

1548-7091

Ausgabe der Veröffentlichung

3

Zeitschrift

Nature Methods

Sprache

en

Online publication date

2011

Paginierung

231 - 237

Datum der Veröffentlichung

2011

Status

Published

Herausgeber

Springer Science and Business Media LLC

Herausgeber URL

http://dx.doi.org/10.1038/nmeth.1561

Datum der Datenerfassung

2023

Titel

A versatile in vivo system for directed dissection of gene expression patterns

Ausgabe der Zeitschrift

8

Datenquelle: Crossref

Abstract

Tissue-specific gene expression using the upstream activating sequence (UAS)–GAL4 binary system has facilitated genetic dissection of many biological processes in Drosophila melanogaster. Refining GAL4 expression patterns or independently manipulating multiple cell populations using additional binary systems are common experimental goals. To simplify these processes, we developed a convertible genetic platform, the integrase swappable in vivo targeting element (InSITE) system. This approach allows GAL4 to be replaced with any other sequence, placing different genetic effectors under the control of the same regulatory elements. Using InSITE, GAL4 can be replaced with LexA or QF, allowing an expression pattern to be repurposed. GAL4 can also be replaced with GAL80 or split-GAL4 hemi-drivers, allowing intersectional approaches to refine expression patterns. The exchanges occur through efficient in vivo manipulations, making it possible to generate many swaps in parallel. This system is modular, allowing future genetic tools to be easily incorporated into the existing framework.

Addresses

Department of Neurobiology, Stanford University, Stanford, California, USA.

Autoren

Daryl M Gohl
Marion A Silies
Xiaojing J Gao
Sheetal Bhalerao
Francisco J Luongo
Chun-Chieh Lin
Christopher J Potter
Thomas R Clandinin

DOI

10.1038/nmeth.1561

eISSN

1548-7105

Externe Identifier

PubMed Identifier: 21473015
PubMed Central ID: PMC3079545

Funding acknowledgements

NIH HHS: DP1 OD003530-05
NEI NIH HHS: R01 EY015231
NIH HHS: DP1 OD003530-01
NIH HHS: DP1 OD003530-02
NIH HHS: DP1 OD003530-04
NIH HHS: DP1 OD003530
NEI NIH HHS: R01 EY015231-07
NIH HHS: DP1 OD003530-03

Open access

true

ISSN

1548-7091

Ausgabe der Veröffentlichung

3

Zeitschrift

Nature methods

Schlüsselwörter

Animals
Drosophila melanogaster
Serine Endopeptidases
Bacterial Proteins
DNA-Binding Proteins
Saccharomyces cerevisiae Proteins
Transcription Factors
Repressor Proteins
Gene Expression Profiling
Gene Expression
Recombination, Genetic
Base Sequence
Molecular Sequence Data
Enhancer Elements, Genetic

Sprache

eng

Medium

Print

Open access status

Open Access

Paginierung

231 - 237

Datum der Veröffentlichung

2011

Status

Published

Datum der Datenerfassung

2011

Titel

A versatile in vivo system for directed dissection of gene expression patterns.

Sub types

Research Support, Non-U.S. Gov't
research-article
Journal Article
Research Support, N.I.H., Extramural

Ausgabe der Zeitschrift

8

Files

https://europepmc.org/articles/PMC3079545?pdf=render

Datenquelle: Europe PubMed Central

Abstract

Tissue-specific gene expression using the upstream activating sequence (UAS)–GAL4 binary system has facilitated genetic dissection of many biological processes in Drosophila melanogaster. Refining GAL4 expression patterns or independently manipulating multiple cell populations using additional binary systems are common experimental goals. To simplify these processes, we developed a convertible genetic platform, the integrase swappable in vivo targeting element (InSITE) system. This approach allows GAL4 to be replaced with any other sequence, placing different genetic effectors under the control of the same regulatory elements. Using InSITE, GAL4 can be replaced with LexA or QF, allowing an expression pattern to be repurposed. GAL4 can also be replaced with GAL80 or split-GAL4 hemi-drivers, allowing intersectional approaches to refine expression patterns. The exchanges occur through efficient in vivo manipulations, making it possible to generate many swaps in parallel. This system is modular, allowing future genetic tools to be easily incorporated into the existing framework.

Autoren

Daryl M Gohl
Marion A Silies
Xiaojing J Gao
Sheetal Bhalerao
Francisco J Luongo
Chun-Chieh Lin
Christopher J Potter
Thomas R Clandinin

Autoren-URL

https://www.ncbi.nlm.nih.gov/pubmed/21473015

DOI

10.1038/nmeth.1561

eISSN

1548-7105

Externe Identifier

NIH Manuscript Submission ID: NIHMS261542
PubMed Central ID: PMC3079545

Funding acknowledgements

NEI NIH HHS: R01 EY015231-07
NIH HHS: DP1 OD003530-01
NIH HHS: DP1 OD003530
NIH HHS: DP1 OD003530-05
NIH HHS: DP1 OD003530-02
NIH HHS: DP1 OD003530-03
NEI NIH HHS: R01 EY015231
NIH HHS: DP1 OD003530-04

Ausgabe der Veröffentlichung

3

Zeitschrift

Nat Methods

Schlüsselwörter

Animals
Bacterial Proteins
Base Sequence
DNA-Binding Proteins
Drosophila melanogaster
Enhancer Elements, Genetic
Gene Expression
Gene Expression Profiling
Molecular Sequence Data
Recombination, Genetic
Repressor Proteins
Saccharomyces cerevisiae Proteins
Serine Endopeptidases
Transcription Factors

Sprache

eng

Country

United States

Paginierung

231 - 237

Datum der Veröffentlichung

2011

Status

Published

Datum, an dem der Datensatz öffentlich gemacht wurde

2011

Titel

A versatile in vivo system for directed dissection of gene expression patterns.

Sub types

Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

Ausgabe der Zeitschrift

8

Datenquelle: PubMed

A versatile in vivo system for directed dissection of gene expression patterns

Files

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