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Publication : Characterization of Cre-mediated cassette exchange after plasmid microinjection in fertilized mouse oocytes.

First Author  Lauth M Year  2000
Journal  Genesis Volume  27
Issue  4 Pages  153-8
PubMed ID  10992325 Mgi Jnum  J:115349
Mgi Id  MGI:3691413 Doi  10.1002/1526-968x(200008)27:4<153::aid-gene40>3.0.co;2-b
Citation  Lauth M, et al. (2000) Characterization of Cre-mediated cassette exchange after plasmid microinjection in fertilized mouse oocytes. Genesis 27(4):153-8
abstractText  DNA cassette exchange methodologies are powerful tools for rapid modification of defined genomic loci. Until now, most studies have focused on transfected cell lines that were subjected to positive and negative selection to enrich for correctly recombined clones. This is the first report on Cre-mediated cassette exchange (CMCE) in early mouse embryos, a nonselectable biological system. A Cre expression plasmid and a replacement plasmid were introduced into fertilized oocytes by plasmid microinjection and recombination products were investigated. Both, replacement plasmid and oocyte genome carried heterospecific lox sites (lox511 and loxP) to direct CMCE toward integration. Here, we demonstrate the general feasibility of CMCE in early mouse embryos and characterize the system with respect to integration efficiency, competing reactions and parameters affecting recombination rates.
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