Rapid screening of gene function by systemic delivery of morpholino oligonucleotides to live mouse embryos.
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 m...
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| Format: | Article |
| Language: | English |
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Public Library of Science (PLoS)
2015-01-01
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| Series: | PLoS ONE |
| Online Access: | https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0114932&type=printable |
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| author | Kathryn S McClelland Elanor N Wainwright Josephine Bowles Peter Koopman |
| author_facet | Kathryn S McClelland Elanor N Wainwright Josephine Bowles Peter Koopman |
| author_sort | Kathryn S McClelland |
| collection | DOAJ |
| description | 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. |
| format | Article |
| id | doaj-art-3c232249f2b34c458fda083639e3cdb1 |
| institution | DOAJ |
| issn | 1932-6203 |
| language | English |
| publishDate | 2015-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj-art-3c232249f2b34c458fda083639e3cdb12025-08-20T03:01:28ZengPublic Library of Science (PLoS)PLoS ONE1932-62032015-01-01101e011493210.1371/journal.pone.0114932Rapid screening of gene function by systemic delivery of morpholino oligonucleotides to live mouse embryos.Kathryn S McClellandElanor N WainwrightJosephine BowlesPeter KoopmanTraditional 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.https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0114932&type=printable |
| spellingShingle | Kathryn S McClelland Elanor N Wainwright Josephine Bowles Peter Koopman Rapid screening of gene function by systemic delivery of morpholino oligonucleotides to live mouse embryos. PLoS ONE |
| title | Rapid screening of gene function by systemic delivery of morpholino oligonucleotides to live mouse embryos. |
| title_full | Rapid screening of gene function by systemic delivery of morpholino oligonucleotides to live mouse embryos. |
| title_fullStr | Rapid screening of gene function by systemic delivery of morpholino oligonucleotides to live mouse embryos. |
| title_full_unstemmed | Rapid screening of gene function by systemic delivery of morpholino oligonucleotides to live mouse embryos. |
| title_short | Rapid screening of gene function by systemic delivery of morpholino oligonucleotides to live mouse embryos. |
| title_sort | rapid screening of gene function by systemic delivery of morpholino oligonucleotides to live mouse embryos |
| url | https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0114932&type=printable |
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