In:
BMC Biotechnology, Springer Science and Business Media LLC, Vol. 8, No. 1 ( 2008-12)
Kurzfassung:
Inducible conditional knockout animals are widely used to get insight in the function of genes and the pathogenesis of human diseases. These models frequently rely on Cre-mediated recombination of sequences flanked by Lox-P sites. To understand the consequences of gene disruption, it is essential to know the efficiency of the recombination process. Results Here, we describe a modification of the multiplex ligation-dependent probe amplification (MLPA), called extension-MLPA (eMLPA), which enables quantification of relatively small differences in DNA that are a consequence of Cre-mediated recombination. eMLPA, here applied on an inducible Pkd1 conditional deletion mouse model, simultaneously measures both the reduction of the floxed allele and the increase of the deletion allele in a single reaction thereby minimizing any type of experimental variation. Interestingly, with this method we were also able to observe the presence of the excised DNA fragment. This extra-chromosomal deletion-circle was detectable up to 5 months after activation of Cre. Conclusion eMLPA is a novel strategy which easily can be applied to measure the Cre-mediated recombination efficiency in each experimental case with high accuracy. In addition the fate of the deletion-circle can be followed simultaneously.
Materialart:
Online-Ressource
ISSN:
1472-6750
DOI:
10.1186/1472-6750-8-18
Sprache:
Englisch
Verlag:
Springer Science and Business Media LLC
Publikationsdatum:
2008
ZDB Id:
2052746-9
SSG:
12
