Genomic integration and ligand-dependent activation of the human estrogen receptor α in the crustacean Daphnia magna

Kerstin Törner; Tsuyoshi Nakanishi; Tomoaki Matsuura; Yasuhiko Kato; Hajime Watanabe

doi:10.1371/journal.pone.0198023

Genomic integration and ligand-dependent activation of the human estrogen receptor α in the crustacean Daphnia magna

PLoS One. 2018 Jun 8;13(6):e0198023. doi: 10.1371/journal.pone.0198023. eCollection 2018.

Authors

Kerstin Törner¹, Tsuyoshi Nakanishi², Tomoaki Matsuura¹, Yasuhiko Kato^{1

3}, Hajime Watanabe¹

Affiliations

¹ Department of Biotechnology, Graduate School of Engineering, Osaka University, Yamadaoka, Suita, Osaka, Japan.
² Laboratory of Hygienic Chemistry and Molecular Toxicology, Gifu Pharmaceutical University, Daigaku-nishi, Gifu, Gifu, Japan.
³ Frontier Research Base for Global Young Researchers, Graduate School of Engineering, Osaka University, Yamadaoka, Suita, Osaka, Japan.

Abstract

The freshwater crustacean Daphnia have a long history in water quality assessments and now lend themselves to detection of targeted chemicals using genetically encoded reporter gene due to recent progress in the development of genome editing tools. By introducing human genes into Daphnia, we may be able to detect chemicals that affect the human system, or even apply it to screening potentially useful chemicals. Here, we aimed to develop a transgenic line of Daphnia magna that contains the human estrogen receptor alpha (hERα) and shows a fluorescence response to exposure of estrogens. We designed plasmids to express hERα in Daphnia (EF1α1:esr1) and to report estrogenic activity via red fluorescence (ERE:mcherry) under the control of estrogen response element (ERE). After confirmation of functionality of the plasmids by microinjection into embryos, the two plasmids were joined, a TALE site was added and integrated into the D. magna genome using TALEN. When the resulting transgenic Daphnia named the ES line was exposed to Diethylstilbestrol (DES) or 17β-Estradiol (E2), the ES line could reliably expressed red fluorescence derived from mCherry in a ligand-dependent manner, indicating that an estrogen-responsive line of D. magna was established. This is the first time a human gene was expressed in Daphnia, showcasing potential for further research.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Animals, Genetically Modified
Daphnia / genetics*
Estrogen Receptor alpha / genetics*
Gene Transfer Techniques*
Genomics*
Ligands

Substances

ESR1 protein, human
Estrogen Receptor alpha
Ligands

Grants and funding

One of the authors (YK) acknowledges the support of the Frontier Research Base for Global Young Researchers, Osaka University. This work was supported by Japan Society for the Promotion of Science KAKENHI Grant Number JP26281027. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.