Conditional RNAi in mice
- PMID: 20705138
- DOI: 10.1016/j.ymeth.2010.08.003
Conditional RNAi in mice
Abstract
RNA interference (RNAi)-mediated gene knockdown has developed into a routine method to assess gene function in cultured mammalian cells in a fast and easy manner. For the use of RNAi in mice, short hairpin (sh) RNAs expressed stably from the genome are a fast alternative to conventional knockout approaches. We developed a strategy for complete or conditional gene knockdown in mice, where the Cre/loxP system is used to activate RNAi in a time and tissue dependent manner. Alternatively doxycycline controlled shRNA expression vectors can be used for conditional gene silencing. Single copy RNAi constructs are placed into the Rosa26 locus of ES cells by recombinase mediated cassette exchange and transmitted through the germline of chimeric mice. The shRNA transgenic offspring can be either directly used for phenotypic analysis or are further crossed to a Cre transgenic strain to activate conditional shRNA vectors. The site specific insertion of single copy shRNA vectors allows the expedite and reproducible production of knockdown mice and provides an easy and fast approach to assess gene function in vivo.
Copyright © 2010 Elsevier Inc. All rights reserved.
Similar articles
-
Generation of shRNA transgenic mice.Methods Mol Biol. 2009;530:101-29. doi: 10.1007/978-1-59745-471-1_6. Methods Mol Biol. 2009. PMID: 19266327
-
Gene knockdown in the mouse through RNAi.Methods Enzymol. 2010;477:387-414. doi: 10.1016/S0076-6879(10)77020-8. Methods Enzymol. 2010. PMID: 20699152
-
Constitutive and conditional RNAi transgenesis in mice.Methods. 2011 Apr;53(4):430-6. doi: 10.1016/j.ymeth.2010.12.015. Epub 2010 Dec 22. Methods. 2011. PMID: 21184828
-
RNA interference in mice.Handb Exp Pharmacol. 2007;(178):149-76. doi: 10.1007/978-3-540-35109-2_7. Handb Exp Pharmacol. 2007. PMID: 17203655 Review.
-
Conditional RNAi: towards a silent gene therapy.Adv Drug Deliv Rev. 2009 Jul 2;61(7-8):650-64. doi: 10.1016/j.addr.2009.03.016. Epub 2009 Apr 24. Adv Drug Deliv Rev. 2009. PMID: 19394374 Review.
Cited by
-
Methods to Investigate miRNA Function: Focus on Platelet Reactivity.Thromb Haemost. 2021 Apr;121(4):409-421. doi: 10.1055/s-0040-1718730. Epub 2020 Oct 29. Thromb Haemost. 2021. PMID: 33124028 Free PMC article. Review.
-
Cell Type-Specific and Inducible PTEN Gene Silencing by a Tetracycline Transcriptional Activator-Regulated Short Hairpin RNA.Mol Cells. 2015 Nov;38(11):959-65. doi: 10.14348/molcells.2015.0137. Epub 2015 Oct 19. Mol Cells. 2015. PMID: 26486163 Free PMC article.
-
Simple and rapid determination of homozygous transgenic mice via in vivo fluorescence imaging.Oncotarget. 2015 Nov 17;6(36):39073-87. doi: 10.18632/oncotarget.5535. Oncotarget. 2015. PMID: 26472024 Free PMC article.
-
Genetically engineered mouse models for drug development and preclinical trials.Biomol Ther (Seoul). 2014 Jul;22(4):267-74. doi: 10.4062/biomolther.2014.074. Biomol Ther (Seoul). 2014. PMID: 25143803 Free PMC article. Review.
-
Fluorescent-based methods for gene knockdown and functional cardiac imaging in zebrafish.Mol Biotechnol. 2013 Oct;55(2):131-42. doi: 10.1007/s12033-013-9664-6. Mol Biotechnol. 2013. PMID: 23674069 Free PMC article.
Publication types
MeSH terms
Substances
LinkOut - more resources
Full Text Sources