A pipeline for the generation of shRNA transgenic mice Journal Article
| Authors: | Dow, L. E.; Premsrirut, P. K.; Zuber, J.; Fellmann, C.; McJunkin, K.; Miething, C.; Park, Y.; Dickins, R. A.; Hannon, G. J.; Lowe, S. W. |
| Article Title: | A pipeline for the generation of shRNA transgenic mice |
| Abstract: | RNA interference (RNAi) is an extremely effective tool for studying gene function in almost all metazoan and eukaryotic model systems. RNAi in mice, through the expression of short hairpin RNAs (shRNAs), offers something not easily achieved with traditional genetic approaches-inducible and reversible gene silencing. However, technical variability associated with the production of shRNA transgenic strains has so far limited their widespread use. Here we describe a pipeline for the generation of miR30-based shRNA transgenic mice that enables efficient and consistent targeting of doxycycline-regulated, fluorescence-linked shRNAs to the Col1a1 locus. Notably, the protocol details crucial steps in the design and testing of miR30-based shRNAs to maximize the potential for developing effective transgenic strains. In all, this 14-week procedure provides a fast and cost-effective way for any laboratory to investigate gene function in vivo in the mouse. © 2012 Nature America, Inc. All rights reserved. |
| Keywords: | controlled study; protein expression; unclassified drug; nonhuman; animal cell; mouse; animals; mice; mus; microrna; protein targeting; animal experiment; rna, small interfering; gene locus; genetic variability; genotype; gene function; rna interference; in vivo study; transgenic mouse; mus musculus; mice, transgenic; collagen type i; cloning, molecular; genetic engineering; eukaryota; gene silencing; micrornas; doxycycline; fluorescence analysis; short hairpin rna; metazoa; mir30; genotyping techniques |
| Journal Title: | Nature Protocols |
| Volume: | 7 |
| Issue: | 2 |
| ISSN: | 1754-2189 |
| Publisher: | Nature Publishing Group |
| Date Published: | 2012-02-01 |
| Start Page: | 374 |
| End Page: | 393 |
| Language: | English |
| DOI: | 10.1038/nprot.2011.446 |
| PROVIDER: | scopus |
| PUBMED: | 22301776 |
| PMCID: | PMC3724521 |
| DOI/URL: | |
| Notes: | --- - "Cited By (since 1996): 2" - "Export Date: 11 July 2012" - "Source: Scopus" |
