A site-specific, single-copy transgenesis strategy to identify 5′ regulatory sequences of the mouse testis- determining gene Sry

Quinn, Alexander, Kashimada, Kenichi, Davidson, Tara-Lynne, Ng, Ee Ting, Chawengsaksophak, Kallayanee, Bowles, Josephine and Koopman, Peter (2014) A site-specific, single-copy transgenesis strategy to identify 5′ regulatory sequences of the mouse testis- determining gene Sry. PLoS One, 9 4: 1-10. doi:10.1371/journal.pone.0094813


Author Quinn, Alexander
Kashimada, Kenichi
Davidson, Tara-Lynne
Ng, Ee Ting
Chawengsaksophak, Kallayanee
Bowles, Josephine
Koopman, Peter
Title A site-specific, single-copy transgenesis strategy to identify 5′ regulatory sequences of the mouse testis- determining gene Sry
Formatted title
A site-specific, single-copy transgenesis strategy to identify 5′ regulatory sequences of the mouse testis- determining gene Sry
Journal name PLoS One   Check publisher's open access policy
ISSN 1932-6203
Publication date 2014-04-17
Year available 2014
Sub-type Article (original research)
DOI 10.1371/journal.pone.0094813
Open Access Status DOI
Volume 9
Issue 4
Start page 1
End page 10
Total pages 10
Place of publication San Francisco, United States
Publisher Public Library of Science
Language eng
Subject 1100 Agricultural and Biological Sciences
1300 Biochemistry, Genetics and Molecular Biology
2700 Medicine
Abstract The Y-chromosomal gene SRY acts as the primary trigger for male sex determination in mammalian embryos. Correct regulation of SRY is critical: aberrant timing or level of Sry expression is known to disrupt testis development in mice and we hypothesize that mutations that affect regulation of human SRY may account for some of the many cases of XY gonadal dysgenesis that currently remain unexplained. However, the cis-sequences involved in regulation of Sry have not been identified, precluding a test of this hypothesis. Here, we used a transgenic mouse approach aimed at identifying mouse Sry 5' flanking regulatory sequences within 8 kb of the Sry transcription start site (TSS). To avoid problems associated with conventional pronuclear injection of transgenes, we used a published strategy designed to yield single-copy transgene integration at a defined, transcriptionally open, autosomal locus, Col1a1. None of the Sry transgenes tested was expressed at levels compatible with activation of Sox9 or XX sex reversal. Our findings indicate either that the Col1a1 locus does not provide an appropriate context for the correct expression of Sry transgenes, or that the cis-sequences required for Sry expression in the developing gonads lie beyond 8 kb 5' of the TSS.
Formatted abstract
The Y-chromosomal gene SRY acts as the primary trigger for male sex determination in mammalian embryos. Correct regulation of SRY is critical: aberrant timing or level of Sry expression is known to disrupt testis development in mice and we hypothesize that mutations that affect regulation of human SRY may account for some of the many cases of XY gonadal dysgenesis that currently remain unexplained. However, the cis-sequences involved in regulation of Sry have not been identified, precluding a test of this hypothesis. Here, we used a transgenic mouse approach aimed at identifying mouse Sry 5′ flanking regulatory sequences within 8 kb of the Sry transcription start site (TSS). To avoid problems associated with conventional pronuclear injection of transgenes, we used a published strategy designed to yield single-copy transgene integration at a defined, transcriptionally open, autosomal locus, Col1a1. None of the Sry transgenes tested was expressed at levels compatible with activation of Sox9 or XX sex reversal. Our findings indicate either that the Col1a1 locus does not provide an appropriate context for the correct expression of Sry transgenes, or that the cis-sequences required for Sry expression in the developing gonads lie beyond 8 kb 5′ of the TSS.
Keyword Multidisciplinary Sciences
Science & Technology - Other Topics
MULTIDISCIPLINARY SCIENCES
Q-Index Code C1
Q-Index Status Confirmed Code
Grant ID DP120104534
546517
Institutional Status UQ
Additional Notes Article # e94813

Document type: Journal Article
Sub-type: Article (original research)
Collections: Official 2015 Collection
Institute for Molecular Bioscience - Publications
 
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