Multiple renal cyst development but not situs abnormalities in transgenic RNAi mice against Inv::GFP rescue gene.
In this study we generated RNA interference (RNAi)-mediated gene knockdown transgenic mice (transgenic RNAi mice) against the functional Inv gene. Inv mutant mice show consistently reversed internal organs (situs inversus), multiple renal cysts and neonatal lethality. The Inv::GFP-rescue mice, which...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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Public Library of Science (PLoS)
2014-01-01
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| Series: | PLoS ONE |
| Online Access: | https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0089652&type=printable |
| _version_ | 1826588628534427648 |
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| author | Yuki Kamijho Yayoi Shiozaki Eiki Sakurai Kazunori Hanaoka Daisuke Watanabe |
| author_facet | Yuki Kamijho Yayoi Shiozaki Eiki Sakurai Kazunori Hanaoka Daisuke Watanabe |
| author_sort | Yuki Kamijho |
| collection | DOAJ |
| description | In this study we generated RNA interference (RNAi)-mediated gene knockdown transgenic mice (transgenic RNAi mice) against the functional Inv gene. Inv mutant mice show consistently reversed internal organs (situs inversus), multiple renal cysts and neonatal lethality. The Inv::GFP-rescue mice, which introduced the Inv::GFP fusion gene, can rescue inv mutant mice phenotypes. This indicates that the Inv::GFP gene is functional in vivo. To analyze the physiological functions of the Inv gene, and to demonstrate the availability of transgenic RNAi mice, we introduced a short hairpin RNA expression vector against GFP mRNA into Inv::GFP-rescue mice and analyzed the gene silencing effects and Inv functions by examining phenotypes. Transgenic RNAi mice with the Inv::GFP-rescue gene (Inv-KD mice) down-regulated Inv::GFP fusion protein and showed hypomorphic phenotypes of inv mutant mice, such as renal cyst development, but not situs abnormalities or postnatal lethality. This indicates that shRNAi-mediated gene silencing systems that target the tag sequence of the fusion gene work properly in vivo, and suggests that a relatively high level of Inv protein is required for kidney development in contrast to left/right axis determination. Inv::GFP protein was significantly down-regulated in the germ cells of Inv-KD mice testis compared with somatic cells, suggesting the existence of a testicular germ cell-specific enhanced RNAi system that regulates germ cell development. The Inv-KD mouse is useful for studying Inv gene functions in adult tissue that are unable to be analyzed in inv mutant mice showing postnatal lethality. In addition, the shRNA-based gene silencing system against the tag sequence of the fusion gene can be utilized as a new technique to regulate gene expression in either in vitro or in vivo experiments. |
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| id | doaj.art-d104302c2f6a4e569cc72a8e768d8455 |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2025-03-14T16:44:21Z |
| publishDate | 2014-01-01 |
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| series | PLoS ONE |
| spelling | doaj.art-d104302c2f6a4e569cc72a8e768d84552025-02-21T05:38:08ZengPublic Library of Science (PLoS)PLoS ONE1932-62032014-01-0192e8965210.1371/journal.pone.0089652Multiple renal cyst development but not situs abnormalities in transgenic RNAi mice against Inv::GFP rescue gene.Yuki KamijhoYayoi ShiozakiEiki SakuraiKazunori HanaokaDaisuke WatanabeIn this study we generated RNA interference (RNAi)-mediated gene knockdown transgenic mice (transgenic RNAi mice) against the functional Inv gene. Inv mutant mice show consistently reversed internal organs (situs inversus), multiple renal cysts and neonatal lethality. The Inv::GFP-rescue mice, which introduced the Inv::GFP fusion gene, can rescue inv mutant mice phenotypes. This indicates that the Inv::GFP gene is functional in vivo. To analyze the physiological functions of the Inv gene, and to demonstrate the availability of transgenic RNAi mice, we introduced a short hairpin RNA expression vector against GFP mRNA into Inv::GFP-rescue mice and analyzed the gene silencing effects and Inv functions by examining phenotypes. Transgenic RNAi mice with the Inv::GFP-rescue gene (Inv-KD mice) down-regulated Inv::GFP fusion protein and showed hypomorphic phenotypes of inv mutant mice, such as renal cyst development, but not situs abnormalities or postnatal lethality. This indicates that shRNAi-mediated gene silencing systems that target the tag sequence of the fusion gene work properly in vivo, and suggests that a relatively high level of Inv protein is required for kidney development in contrast to left/right axis determination. Inv::GFP protein was significantly down-regulated in the germ cells of Inv-KD mice testis compared with somatic cells, suggesting the existence of a testicular germ cell-specific enhanced RNAi system that regulates germ cell development. The Inv-KD mouse is useful for studying Inv gene functions in adult tissue that are unable to be analyzed in inv mutant mice showing postnatal lethality. In addition, the shRNA-based gene silencing system against the tag sequence of the fusion gene can be utilized as a new technique to regulate gene expression in either in vitro or in vivo experiments.https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0089652&type=printable |
| spellingShingle | Yuki Kamijho Yayoi Shiozaki Eiki Sakurai Kazunori Hanaoka Daisuke Watanabe Multiple renal cyst development but not situs abnormalities in transgenic RNAi mice against Inv::GFP rescue gene. PLoS ONE |
| title | Multiple renal cyst development but not situs abnormalities in transgenic RNAi mice against Inv::GFP rescue gene. |
| title_full | Multiple renal cyst development but not situs abnormalities in transgenic RNAi mice against Inv::GFP rescue gene. |
| title_fullStr | Multiple renal cyst development but not situs abnormalities in transgenic RNAi mice against Inv::GFP rescue gene. |
| title_full_unstemmed | Multiple renal cyst development but not situs abnormalities in transgenic RNAi mice against Inv::GFP rescue gene. |
| title_short | Multiple renal cyst development but not situs abnormalities in transgenic RNAi mice against Inv::GFP rescue gene. |
| title_sort | multiple renal cyst development but not situs abnormalities in transgenic rnai mice against inv gfp rescue gene |
| url | https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0089652&type=printable |
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