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Rapid and Efficient Generation of Recombinant Human Pluripotent Stem Cells by Recombinase-mediated Cassette Exchange in the AAVS1 Locus
Published on: November 20, 2016
P22 mediated recombination of frt-sites
Angela Makumi1, William Cenens1, Rob Lavigne2
1Laboratory of Food Microbiology, Department of Microbial and Molecular Systems (M(2)S), Faculty of Bioscience Engineering, KU Leuven, Kasteelpark Arenberg 23, B-3001 Leuven, Belgium.
Abstract:
Flp mediated site specific recombination of frt-sites is frequently used in genetic engineering to excise, insert or invert DNA-cassettes in the chromosome. While constructs flanked by frt-sites are generally considered to be stable in the absence of the Flp enzyme, we observed that P22 chromosomes exceeding wild-type length tend to lose frt-flanked insertions via Flp independent recombination of frt-sites during phage propagation. This spontaneous recombination should be considered when engineering the chromosome of P22 and perhaps of other phages as well.
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