Multiple renal cyst development but not situs abnormalities in transgenic RNAi mice against Inv::GFP rescue gene
| Autor: | Eiki Sakurai, Yuki Kamijho, Daisuke Watanabe, Kazunori Hanaoka, Yayoi Shiozaki |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2014 |
| Předmět: |
Male
Embryology Aging Mouse Mutant lcsh:Medicine Kidney Small hairpin RNA Molecular cell biology RNA interference Testis Gene Knockdown Techniques Morphogenesis Pattern Formation RNA Small Interfering lcsh:Science Animal Management Gene knockdown Multidisciplinary Expression vector Stem Cells Agriculture Cell Differentiation Genomics Animal Models Kidney Diseases Cystic Situs Inversus Gene Fusion Genetic Engineering Transgenic Animals Research Article Biotechnology Histology Transgene Green Fluorescent Proteins Bioengineering Mice Transgenic Biology Model Organisms Genome Analysis Tools Gene silencing Animals Gene Silencing fungi lcsh:R Body Plan Organization Molecular Development biochemical phenomena metabolism and nutrition Molecular biology Small Molecules lcsh:Q Gene expression Organism Development Transgenics Developmental Biology Transcription Factors |
| Zdroj: | PLoS ONE, Vol 9, Iss 2, p e89652 (2014) PLoS ONE |
| ISSN: | 1932-6203 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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