Tol2 transposon-mediated transgenesis in the Midas cichlid (Amphilophus citrinellus) : towards understanding gene function and regulatory evolution in an ecological model system for rapid phenotypic diversification

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Kratochwil_2-izpa0crd9ydn8.pdf
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2017
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Sefton, Maggie M.
Liang, Yipeng
Meyer, Axel
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European Union (EU): 293700
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Comparative genomics of parallel in repeated adaptive radiations (GenAdap)
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BMC Developmental Biology. 2017, 17, 15. eISSN 1471-213X. Available under: doi: 10.1186/s12861-017-0157-x
Zusammenfassung

Background
The Midas cichlid species complex (Amphilophus spp.) is widely known among evolutionary biologists as a model system for sympatric speciation and adaptive phenotypic divergence within extremely short periods of time (a few hundred generations). The repeated parallel evolution of adaptive phenotypes in this radiation, combined with their near genetic identity, makes them an excellent model for studying phenotypic diversification. While many ecological and evolutionary studies have been performed on Midas cichlids, the molecular basis of specific phenotypes, particularly adaptations, and their underlying coding and cis-regulatory changes have not yet been studied thoroughly.

Results
For the first time in any New World cichlid, we use Tol2 transposon-mediated transgenesis in the Midas cichlid (Amphilophus citrinellus). By adapting existing microinjection protocols, we established an effective protocol for transgenesis in Midas cichlids. Embryos were injected with a Tol2 plasmid construct that drives enhanced green fluorescent protein (eGFP) expression under the control of the ubiquitin promoter. The transgene was successfully integrated into the germline, driving strong ubiquitous expression of eGFP in the first transgenic Midas cichlid line. Additionally, we show transient expression of two further transgenic constructs, ubiquitin::tdTomato and mitfa::eGFP. Transgenesis in Midas cichlids will facilitate further investigation of the genetic basis of species-specific traits, many of which are adaptations.

Conclusion
Transgenesis is a versatile tool not only for studying regulatory elements such as promoters and enhancers, but also for testing gene function through overexpression of allelic gene variants. As such, it is an important first step in establishing the Midas cichlid as a powerful model for studying adaptive coding and non-coding changes in an ecological and evolutionary context.

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Fachgebiet (DDC)
570 Biowissenschaften, Biologie
Schlagwörter
Transgenesis, Gene function, Transcriptional regulation, Cichlidae, Amphilophus citrinellus, Adaptive radiation
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ISO 690KRATOCHWIL, Claudius F., Maggie M. SEFTON, Yipeng LIANG, Axel MEYER, 2017. Tol2 transposon-mediated transgenesis in the Midas cichlid (Amphilophus citrinellus) : towards understanding gene function and regulatory evolution in an ecological model system for rapid phenotypic diversification. In: BMC Developmental Biology. 2017, 17, 15. eISSN 1471-213X. Available under: doi: 10.1186/s12861-017-0157-x
BibTex
@article{Kratochwil2017-11-23trans-40887,
  year={2017},
  doi={10.1186/s12861-017-0157-x},
  title={Tol2 transposon-mediated transgenesis in the Midas cichlid (Amphilophus citrinellus) : towards understanding gene function and regulatory evolution in an ecological model system for rapid phenotypic diversification},
  volume={17},
  journal={BMC Developmental Biology},
  author={Kratochwil, Claudius F. and Sefton, Maggie M. and Liang, Yipeng and Meyer, Axel},
  note={Article Number: 15}
}
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    <dcterms:abstract xml:lang="eng">Background&lt;br /&gt;The Midas cichlid species complex (Amphilophus spp.) is widely known among evolutionary biologists as a model system for sympatric speciation and adaptive phenotypic divergence within extremely short periods of time (a few hundred generations). The repeated parallel evolution of adaptive phenotypes in this radiation, combined with their near genetic identity, makes them an excellent model for studying phenotypic diversification. While many ecological and evolutionary studies have been performed on Midas cichlids, the molecular basis of specific phenotypes, particularly adaptations, and their underlying coding and cis-regulatory changes have not yet been studied thoroughly.&lt;br /&gt;&lt;br /&gt;Results&lt;br /&gt;For the first time in any New World cichlid, we use Tol2 transposon-mediated transgenesis in the Midas cichlid (Amphilophus citrinellus). By adapting existing microinjection protocols, we established an effective protocol for transgenesis in Midas cichlids. Embryos were injected with a Tol2 plasmid construct that drives enhanced green fluorescent protein (eGFP) expression under the control of the ubiquitin promoter. The transgene was successfully integrated into the germline, driving strong ubiquitous expression of eGFP in the first transgenic Midas cichlid line. Additionally, we show transient expression of two further transgenic constructs, ubiquitin::tdTomato and mitfa::eGFP. Transgenesis in Midas cichlids will facilitate further investigation of the genetic basis of species-specific traits, many of which are adaptations.&lt;br /&gt;&lt;br /&gt;Conclusion&lt;br /&gt;Transgenesis is a versatile tool not only for studying regulatory elements such as promoters and enhancers, but also for testing gene function through overexpression of allelic gene variants. As such, it is an important first step in establishing the Midas cichlid as a powerful model for studying adaptive coding and non-coding changes in an ecological and evolutionary context.</dcterms:abstract>
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