Rapid screening of gene function by systemic delivery of morpholino oligonucleotides to live mouse embryos

McClelland, Kathryn S., Wainwright, Elanor N., Bowles, Josephine and Koopman, Peter (2015) Rapid screening of gene function by systemic delivery of morpholino oligonucleotides to live mouse embryos. PLoS One, 10 1: 1-18. doi:10.1371/journal.pone.0114932


Author McClelland, Kathryn S.
Wainwright, Elanor N.
Bowles, Josephine
Koopman, Peter
Title Rapid screening of gene function by systemic delivery of morpholino oligonucleotides to live mouse embryos
Journal name PLoS One   Check publisher's open access policy
ISSN 1932-6203
1932-6203
Publication date 2015-01-28
Year available 2015
Sub-type Article (original research)
DOI 10.1371/journal.pone.0114932
Open Access Status DOI
Volume 10
Issue 1
Start page 1
End page 18
Total pages 18
Place of publication San Francisco CA, United States
Publisher Public Library of Science
Language eng
Formatted abstract
Traditional gene targeting methods in mice are complex and time consuming, especially when conditional deletion methods are required. Here, we describe a novel technique for assessing gene function by injection of modified antisense morpholino oligonucleotides (MOs) into the heart of mid-gestation mouse embryos. After allowing MOs to circulate through the embryonic vasculature, target tissues were explanted, cultured and analysed for expression of key markers. We established proof-of-principle by partially phenocopying known gene knockout phenotypes in the fetal gonads (Stra8, Sox9) and pancreas (Sox9). We also generated a novel double knockdown of Gli1 and Gli2, revealing defects in Leydig cell differentiation in the fetal testis. Finally, we gained insight into the roles of Adamts19 and Ctrb1, genes of unknown function in sex determination and gonadal development. These studies reveal the utility of this method as a means of first-pass analysis of gene function during organogenesis before committing to detailed genetic analysis.
Keyword Premature ovarian failure
Branching morphogenesis
Testis development
Kidney development
Sex determination
Vivo morpholinos
Cell fate
In vivo
Sox9
Expression
Q-Index Code C1
Q-Index Status Confirmed Code
Institutional Status UQ

Document type: Journal Article
Sub-type: Article (original research)
Collections: Official 2016 Collection
Institute for Molecular Bioscience - Publications
 
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Citation counts: TR Web of Science Citation Count  Cited 5 times in Thomson Reuters Web of Science Article | Citations
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Created: Sat, 27 Jun 2015, 01:10:47 EST by Susan Allen on behalf of Institute for Molecular Bioscience