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Publication : Gene targeting of ErbB3 using a Cre-mediated unidirectional DNA inversion strategy.

First Author  Qu S Year  2006
Journal  Genesis Volume  44
Issue  10 Pages  477-86
PubMed ID  16991114 Mgi Jnum  J:114524
Mgi Id  MGI:3689267 Doi  10.1002/dvg.20243
Citation  Qu S, et al. (2006) Gene targeting of ErbB3 using a Cre-mediated unidirectional DNA inversion strategy. Genesis 44(10):477-86
abstractText  Recombinase-mediated unidirectional DNA inversion and transcriptional arrest is a promising strategy for high throughput conditional mutagenesis in the mouse. Banks of mouse embryonic stem cells with defined, transcriptionally silent insertions that can be activated by Cre recombinase would take advantage of existing transgenic Cre lines to rapidly produce hundreds of lineage specific and temporally controlled knockout mice for each gene, thereby introducing significant parallelism to functional gene annotation. However, the extent to which this strategy results in effective gene knockout has not been established. To test the feasibility of this strategy we targeted ErbB3, a member of the ErbB family of tyrosine kinase receptors, using this strategy. Insertion of a reversed 'flipflox' vector consisting of a gene inactivation cassette (GI) and an internal ribosome entry site (IRES)-GFP reporter into intron 1 of ErbB3 was transcriptionally silent and did not affect ErbB3 expression. Crosses with ubiquitous and lineage specific Cre recombinase expressing lines permanently inverted the inserted GI cassette and blocked ErbB3 expression. Unidirectional DNA inversion by in vivo recombination is an effective strategy for targeted or ubiquitous gene knockout. genesis 44:477-486, 2006. Published 2006 Wiley-Liss, Inc.
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