piggyBac-based insertional mutagenesis and enhancer detection as a tool for functional insect genomics
(2003) In Genetics 163(2). p.647-661- Abstract
- Transposon mutagenesis provides a fundamental tool for functional genomics. Here we present a non-species-specific, combined enhancer detection and binary expression system based on the transposable element piggyBac: For the different components of this insertional mutagenesis system, we used widely applicable transposons and distinguishable broad-range transformation markers, which should enable this system to be operational in nonmodel arthropods. In a pilot screen in Drosophila melanogaster, piggyBac mutator elements on the X chromosome were mobilized in males by a Hennes-based jumpstarter element providing pigpBac transposase activity under control of the alpha1-tubulin promoter. As primary reporters in the pigoBac mutator elements, we... (More)
- Transposon mutagenesis provides a fundamental tool for functional genomics. Here we present a non-species-specific, combined enhancer detection and binary expression system based on the transposable element piggyBac: For the different components of this insertional mutagenesis system, we used widely applicable transposons and distinguishable broad-range transformation markers, which should enable this system to be operational in nonmodel arthropods. In a pilot screen in Drosophila melanogaster, piggyBac mutator elements on the X chromosome were mobilized in males by a Hennes-based jumpstarter element providing pigpBac transposase activity under control of the alpha1-tubulin promoter. As primary reporters in the pigoBac mutator elements, we employed the heterologous transactivators GAL4Delta or tTA. To identify larval and adult enhancer detectors, strains carrying UASp-EYFP or TRE-EYFP as secondary reporter elements were used. Tissue-specific enhancer activities were readily observed in the GAL4Delta/UASp-based systems, but only rarely in the tTA/TRE system. Novel autosomal insertions were recovered with an average jumping rate of 80%. Of these novel insertions, 3.8% showed homozygous lethality, which was reversible by pigoBac excision. Insertions were found in both coding and noncoding regions of characterized genes and also in noncharacterized and non-P-targeted CG-number genes. This indicates that piggyBac will greatly facilitate the intended saturation mutagenesis in Drosophila. (Less)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/316987
- author
- Horn, C ; Offen, N ; Nystedt, Sverker LU ; Häcker, Udo LU and Wimmer, EA
- organization
- publishing date
- 2003
- type
- Contribution to journal
- publication status
- published
- subject
- in
- Genetics
- volume
- 163
- issue
- 2
- pages
- 647 - 661
- publisher
- Genetics Society of America
- external identifiers
-
- wos:000181417200019
- pmid:12618403
- scopus:0037295418
- ISSN
- 0016-6731
- language
- English
- LU publication?
- yes
- id
- 4395defb-fbdf-4863-b4e6-8024e4552a3d (old id 316987)
- alternative location
- http://www.genetics.org/cgi/content/abstract/163/2/647
- date added to LUP
- 2016-04-01 11:34:06
- date last changed
- 2022-03-12 21:39:12
@article{4395defb-fbdf-4863-b4e6-8024e4552a3d,
abstract = {{Transposon mutagenesis provides a fundamental tool for functional genomics. Here we present a non-species-specific, combined enhancer detection and binary expression system based on the transposable element piggyBac: For the different components of this insertional mutagenesis system, we used widely applicable transposons and distinguishable broad-range transformation markers, which should enable this system to be operational in nonmodel arthropods. In a pilot screen in Drosophila melanogaster, piggyBac mutator elements on the X chromosome were mobilized in males by a Hennes-based jumpstarter element providing pigpBac transposase activity under control of the alpha1-tubulin promoter. As primary reporters in the pigoBac mutator elements, we employed the heterologous transactivators GAL4Delta or tTA. To identify larval and adult enhancer detectors, strains carrying UASp-EYFP or TRE-EYFP as secondary reporter elements were used. Tissue-specific enhancer activities were readily observed in the GAL4Delta/UASp-based systems, but only rarely in the tTA/TRE system. Novel autosomal insertions were recovered with an average jumping rate of 80%. Of these novel insertions, 3.8% showed homozygous lethality, which was reversible by pigoBac excision. Insertions were found in both coding and noncoding regions of characterized genes and also in noncharacterized and non-P-targeted CG-number genes. This indicates that piggyBac will greatly facilitate the intended saturation mutagenesis in Drosophila.}},
author = {{Horn, C and Offen, N and Nystedt, Sverker and Häcker, Udo and Wimmer, EA}},
issn = {{0016-6731}},
language = {{eng}},
number = {{2}},
pages = {{647--661}},
publisher = {{Genetics Society of America}},
series = {{Genetics}},
title = {{piggyBac-based insertional mutagenesis and enhancer detection as a tool for functional insect genomics}},
url = {{http://www.genetics.org/cgi/content/abstract/163/2/647}},
volume = {{163}},
year = {{2003}},
}