Development of Cre-loxP technology in zebrafish to study the regulation of fish reproduction

Heng Ju Lin; Shu Hua Lee; Jen Leih Wu; Yeh Fang Duann; Jyh Yih Chen

doi:10.1007/s10695-013-9806-6

Development of Cre-loxP technology in zebrafish to study the regulation of fish reproduction

Heng Ju Lin
, Shu Hua Lee
, Jen Leih Wu
, Yeh Fang Duann
, Jyh Yih Chen

Department of Chemical Engineering and Biotechnology

Research output: Contribution to journal › Article › peer-review

12 Scopus citations

Abstract

One cannot seek permission to market transgenic fish mainly because there is no field test or any basic research on technological developments for evaluating their biosafety. Infertility is a necessary adjunct to exploiting transgenic fish unless completely secure land-locked facilities are available. In this study, we report the generation of a Cre transgenic zebrafish line using a cytomegalovirus promoter. We also produced fish carrying the Bax1 and Bax2 plasmids; these genes were separated by two loxP sites under a zona pellucida C promoter or were driven by an anti-Müllerian hormone promoter. We inserted a red fluorescent protein gene between the two loxP sites. After obtaining transgenic lines with the two transgenic fish crossed with each other (Cre transgenic zebrafish x loxP transgenic zebrafish), the floxed DNA was found to be specifically eliminated from the female or male zebrafish, and apoptosis gene expressions caused ovarian and testicular growth cessation and degeneration. Overexpression of the Bax1 and Bax2 genes caused various expression levels of apoptosis-related genes. Accordingly, this transgenic zebrafish model system provides a method to produce infertile fish and may be useful for application to genetically modified fish.

Original language	English
Pages (from-to)	1525-1539
Number of pages	15
Journal	Fish Physiology and Biochemistry
Volume	39
Issue number	6
DOIs	https://doi.org/10.1007/s10695-013-9806-6
State	Published - Dec 2013

Bibliographical note

Funding Information:
Acknowledgments This work was supported by a grant from the Development Program of Industrialization for Agricultural Biotechnology (money provided by Academia Sinica), called ‘‘Study and development of novel pattern of transgenic fluorescent ornamental fish and infertility technology’’ to Dr. Jyh-Yih Chen in 2010 and 2011. The Cre–loxP plasmid was kindly provided as a gift from Dr. Allan Bradley (Wellcome Trust Sanger Institute, Cambridge, UK).

Keywords

Cre-loxP technology
Infertility
Transgene fish
Zebrafish

Access to Document

10.1007/s10695-013-9806-6

Cite this

@article{e4e444d136dd4424ae3c149f8fa3c7b2,

title = "Development of Cre-loxP technology in zebrafish to study the regulation of fish reproduction",

abstract = "One cannot seek permission to market transgenic fish mainly because there is no field test or any basic research on technological developments for evaluating their biosafety. Infertility is a necessary adjunct to exploiting transgenic fish unless completely secure land-locked facilities are available. In this study, we report the generation of a Cre transgenic zebrafish line using a cytomegalovirus promoter. We also produced fish carrying the Bax1 and Bax2 plasmids; these genes were separated by two loxP sites under a zona pellucida C promoter or were driven by an anti-M{\"u}llerian hormone promoter. We inserted a red fluorescent protein gene between the two loxP sites. After obtaining transgenic lines with the two transgenic fish crossed with each other (Cre transgenic zebrafish x loxP transgenic zebrafish), the floxed DNA was found to be specifically eliminated from the female or male zebrafish, and apoptosis gene expressions caused ovarian and testicular growth cessation and degeneration. Overexpression of the Bax1 and Bax2 genes caused various expression levels of apoptosis-related genes. Accordingly, this transgenic zebrafish model system provides a method to produce infertile fish and may be useful for application to genetically modified fish.",

keywords = "Cre-loxP technology, Infertility, Transgene fish, Zebrafish",

author = "Lin, \{Heng Ju\} and Lee, \{Shu Hua\} and Wu, \{Jen Leih\} and Duann, \{Yeh Fang\} and Chen, \{Jyh Yih\}",

note = "Funding Information: Acknowledgments This work was supported by a grant from the Development Program of Industrialization for Agricultural Biotechnology (money provided by Academia Sinica), called {\textquoteleft}{\textquoteleft}Study and development of novel pattern of transgenic fluorescent ornamental fish and infertility technology{\textquoteright}{\textquoteright} to Dr. Jyh-Yih Chen in 2010 and 2011. The Cre–loxP plasmid was kindly provided as a gift from Dr. Allan Bradley (Wellcome Trust Sanger Institute, Cambridge, UK).",

year = "2013",

month = dec,

doi = "10.1007/s10695-013-9806-6",

language = "???core.languages.en\_GB???",

volume = "39",

pages = "1525--1539",

journal = "Fish Physiology and Biochemistry",

issn = "0920-1742",

publisher = "Springer Science and Business Media B.V.",

number = "6",

}

TY - JOUR

T1 - Development of Cre-loxP technology in zebrafish to study the regulation of fish reproduction

AU - Lin, Heng Ju

AU - Lee, Shu Hua

AU - Wu, Jen Leih

AU - Duann, Yeh Fang

AU - Chen, Jyh Yih

N1 - Funding Information: Acknowledgments This work was supported by a grant from the Development Program of Industrialization for Agricultural Biotechnology (money provided by Academia Sinica), called ‘‘Study and development of novel pattern of transgenic fluorescent ornamental fish and infertility technology’’ to Dr. Jyh-Yih Chen in 2010 and 2011. The Cre–loxP plasmid was kindly provided as a gift from Dr. Allan Bradley (Wellcome Trust Sanger Institute, Cambridge, UK).

PY - 2013/12

Y1 - 2013/12

N2 - One cannot seek permission to market transgenic fish mainly because there is no field test or any basic research on technological developments for evaluating their biosafety. Infertility is a necessary adjunct to exploiting transgenic fish unless completely secure land-locked facilities are available. In this study, we report the generation of a Cre transgenic zebrafish line using a cytomegalovirus promoter. We also produced fish carrying the Bax1 and Bax2 plasmids; these genes were separated by two loxP sites under a zona pellucida C promoter or were driven by an anti-Müllerian hormone promoter. We inserted a red fluorescent protein gene between the two loxP sites. After obtaining transgenic lines with the two transgenic fish crossed with each other (Cre transgenic zebrafish x loxP transgenic zebrafish), the floxed DNA was found to be specifically eliminated from the female or male zebrafish, and apoptosis gene expressions caused ovarian and testicular growth cessation and degeneration. Overexpression of the Bax1 and Bax2 genes caused various expression levels of apoptosis-related genes. Accordingly, this transgenic zebrafish model system provides a method to produce infertile fish and may be useful for application to genetically modified fish.

AB - One cannot seek permission to market transgenic fish mainly because there is no field test or any basic research on technological developments for evaluating their biosafety. Infertility is a necessary adjunct to exploiting transgenic fish unless completely secure land-locked facilities are available. In this study, we report the generation of a Cre transgenic zebrafish line using a cytomegalovirus promoter. We also produced fish carrying the Bax1 and Bax2 plasmids; these genes were separated by two loxP sites under a zona pellucida C promoter or were driven by an anti-Müllerian hormone promoter. We inserted a red fluorescent protein gene between the two loxP sites. After obtaining transgenic lines with the two transgenic fish crossed with each other (Cre transgenic zebrafish x loxP transgenic zebrafish), the floxed DNA was found to be specifically eliminated from the female or male zebrafish, and apoptosis gene expressions caused ovarian and testicular growth cessation and degeneration. Overexpression of the Bax1 and Bax2 genes caused various expression levels of apoptosis-related genes. Accordingly, this transgenic zebrafish model system provides a method to produce infertile fish and may be useful for application to genetically modified fish.

KW - Cre-loxP technology

KW - Infertility

KW - Transgene fish

KW - Zebrafish

UR - https://www.scopus.com/pages/publications/84888005292

U2 - 10.1007/s10695-013-9806-6

DO - 10.1007/s10695-013-9806-6

M3 - ???researchoutput.researchoutputtypes.contributiontojournal.article???

C2 - 23670400

AN - SCOPUS:84888005292

SN - 0920-1742

VL - 39

SP - 1525

EP - 1539

JO - Fish Physiology and Biochemistry

JF - Fish Physiology and Biochemistry

IS - 6

ER -

Development of Cre-loxP technology in zebrafish to study the regulation of fish reproduction

Abstract

Bibliographical note

Keywords

Access to Document

Other files and links

Fingerprint

Cite this