Journal
GENESIS
Volume 50, Issue 6, Pages 482-489Publisher
WILEY
DOI: 10.1002/dvg.20826
Keywords
Cre deleter; Flp deleter; eGFP; eYFP; T2A fusion; targeted transgenesis
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Funding
- CREATE Consortium from the European Commission [223487]
- EUCOMMTOOLS Consortium from the European Commission [223487]
- EUCOMMTOOLS [261492]
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To facilitate the use of the new mutant resource developed in the mouse, we have generated Cre and FlpO deleter mice on a pure inbred C57BL/6N background. The new transgenic constructs were designed to drive either the Cre or FlpO recombinase, fused to a specific fluorescent marker, respectively the eGFP or the eYFP, and were inserted by homologous recombination in the neutral Rosa26 locus. They allow a rapid, cost-effective, and efficient identification of the carrier individuals through the coexpression of the fluorescent marker. The recombination efficiency of the two deleter lines, Gt(ROSA)26Sor < tm1(ACTB-cre,-EGFP)Ics > and Gt(ROSA)26Sor < tm2(CAG-flpo, EYFP)Ics >, was carefully evaluated using five loxP-flanked or four FRT-flanked alleles located at different positions in the mouse genome. For each tested locus, we observed a 100% excision rate. The transgenic mice are easily distinguishable from wild type animals by their bright fluorescence that remains easily detectable until 10 days after birth. In the adult, fluorescence can still be detected inthe unpigmented paws. Furthermore, they both display accumulation of the specific recombinase during oogenesis. These fluorescent Cre- and Flp- deleter transgenic lines are valuable tools for the scientific community by their high and stable recombination efficiency, the simplicity of genotype identification and the maintenance of a pure genetic background when used to remove specific selection cassette or to induce complete loss-of-function allele. genesis 50:482489, 2012. (C) 2012 Wiley Periodicals, Inc.
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