A Transient Transgenic RNAi Strategy for Rapid Characterization of Gene Function during Embryonic Development
Year: 2010
Type: article
Abstract: RNA interference (RNAi) is a powerful strategy for studying the phenotypic consequences of reduced gene expression levels in model systems. To develop a method for the rapid characterization of the developmental consequences of gene dysregulation, we tested the use of RNAi for "transient transgenic" knockdown of mRNA in mouse embryos. These methods included lentiviral infection as well as transposition using the Sleeping Beauty (SB) and PiggyBac (PB) transposable element systems. This approach can be useful for phenotypic validation of putative mutant loci, as we demonstrate by confirming that knockdown of Prdm16 phenocopies the ENU-induced cleft palate (CP) mutant, csp1. This strategy is attractive as an alternative to gene targeting in embryonic stem cells, as it is simple and yields phenotypic information in a matter of weeks. Of the three methodologies tested, the PB transposon system produced high numbers of transgenic embryos with the expected phenotype, demonstrating its utility as a screening method.
Source: PLoS ONE
Language: English
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: gold
Estimated APC paid: $2,382
Funders: Michigan Economic Development Corporation, University of Michigan, National Institutes of Health, Michigan Technology Tri-Corridor, National Institute on Aging, National Institute of Mental Health, National Cancer Institute, National Institute of Diabetes and Digestive and Kidney Diseases, National Institute of Arthritis and Musculoskeletal and Skin Diseases, National Institute of Dental and Craniofacial Research, Eunice Kennedy Shriver National Institute of Child Health and Human Development
Awards: University of Michigan Center for Gastrointestinal Research, Harvard-SDM/Forsyth Scholar/Faculty Development Program, Characterization of the Mouse cpo1 Mutation, ARTHRITIS AND MUSCULOSKELETAL DISEASES CENTER, G3711721818, G4067672505, G4111079442, A Panhypopitutitary Mouse Mutation, Cell specific expression in the pituitary gland, G4840808658, MUTAGENESIS AND MURINE EMBRYONIC DEVELOPMENT, G6712964144, Analysis of neurodevelopment using mouse mutagenesis, Cellular and Molecular Biology of Aging, G818111521, G9000163722, G9000206347, G9000265101, G9000270911, University of Michigan Rogel Cancer Center Support Grant 2023-2028
OpenAlex ID: W2161063308