کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
4156952 1273784 2011 9 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
A more efficient method to generate null mutants using Hprt-Cre with floxed alleles
موضوعات مرتبط
علوم پزشکی و سلامت پزشکی و دندانپزشکی پریناتولوژی (پزشکی مادر و جنین)، طب اطفال و بهداشت کودک
پیش نمایش صفحه اول مقاله
A more efficient method to generate null mutants using Hprt-Cre with floxed alleles
چکیده انگلیسی

PurposeThe generation of nonviable homozygous null mouse embryos from heterozygote null/+ breedings can be highly resource consuming, with only 25% of the embryos in the litter being null mutants. We hypothesized that (1) we could double the number of homozygous null mouse embryos in a litter without reducing litter size using Hypoxanthine-guanine phosphoribosyltransferase-Cre (Hprt)-Cre (which is active in the female germ line at the time of fertilization), and (2) these homozygous null mutants would be identical to mutants generated through traditional null/+ breedings.MethodsTo test this hypothesis, we used a conditional allele Fgfr2IIIbflox. This allele when recombined is identical to the Fgfr2IIIbnull allele. An F1 generation of Fgfr2IIIbrec/+; HprtCre/+ females was created by mating Fgfr2IIIb+/+; Hprtcre/cre females to a Fgfr2IIIbflox/flox male. The F1 females were then mated to a Fgfr2IIIbflox/flox male. F2 embryos were genotyped, and the morphology and histology of the lungs, intestine, limbs, and brain were analyzed.ResultsThe Hprt-Cre mating strategy results in 51% of pups being genotypic homozygous null embryos (85/166) vs 23% for the standard null/+ approach (38/167). These embryos did not express the Fgfr2IIIb transcript and were phenotypically identical to null embryos generated through standard null/+ breedings.ConclusionsThe Hprt-Cre mating strategy increases the number of homozygous mutant embryos in a litter without decreasing litter size. Embryos generated through this approach are phenotypically identical to those from standard heterozygous breedings. We recommend this approach to investigators using a model system that relies on the generation of homozygous null embryos.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Journal of Pediatric Surgery - Volume 46, Issue 9, September 2011, Pages 1711–1719
نویسندگان
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