The promiscuity of heterospecific lox sites increases dramatically in the presence of palindromic DNA.
Heterospecific lox sites are mutated lox sites that in the presence of Cre recombinase recombine with themselves but not or much less with wildtype loxP. We here show that in Escherichia coli both lox511 and lox2272 sites become highly promiscuous with respect to loxP when in the presence of Cre one of the recombination partners is present in a larger stretch of an inverted repeat of non-lox DNA. In such a palindromic DNA configuration, also the occurrence of other DNA repeat-mediated recombination events is somewhat increased in the presence of Cre. The results indicate that in recombinase mediated cassette exchange or other double lox applications based on the exclusivity of heterospecific lox sites, or in research combining Cre-lox approaches with hairpin RNA for gene silencing, the presence of duplicated DNA around lox sites has to be taken into account. It is proposed that the presence of palindromic non-lox DNA interferes with the homology search of the Cre enzyme prior to the actual recombination event.
Mlynárová
Ludmila
L
Plant Research International, Wageningen University and Research Centre, The Netherlands.
Libantová
Jana
J
Vrba
Lukas
L
Nap
Jan-Peter
JP
D001665:Binding Sites / Q000235:genetics; D004269:DNA, Bacterial / Q000235:genetics / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics; D019426:Integrases / Q000235:genetics / Q000378:metabolism*; D008957:Models, Genetic; D009154:Mutation; D010957:Plasmids / Q000235:genetics; D011995:Recombination, Genetic / Q000235:genetics*; D012091:Repetitive Sequences, Nucleic Acid / Q000235:genetics*; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism*
D004269:DNA, Bacterial; D014764:Viral Proteins; C045073:Cre recombinase; D019426:Integrases