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Fig. 1. Cardiac-specific Cre-mediated recombination of Hand1. (A) LoxP
sites were inserted into the 5'-UTR and intron to flank the first exon
of Hand1. The structure of the Hand1 genomic locus, the
targeting vector and the targeted allele are shown. The neomycin resistance
cassette was removed in the mouse germline by breeding heterozygous mice to
hACTB::FLPe transgenic mice. Cardiac-specific excision of exon 1 was
achieved by breeding Hand1loxP/loxP mice to
Hand1LacZ/+ heterozygous mice harboring a transgene that
expresses Cre under the control of 
MHC promoter or
Nkx2.5 cardiac enhancer. Positions of the probe used for Southern
analysis and the primers (a and b) used for RT-PCR are shown. Coding regions
are shown in white and non-coding regions are shown in gray. (B) Detection of
all four Hand1 alleles by Southern blot analysis of
EcoRI-digested genomic DNA, using the 3' probe shown in A. (C)
MHC::Cre transgenic mice were intercrossed to ROSA26R
indicator mice to determine the temporal and tissue specificity of
recombination. Whole-mount photographs of ß-gal stained embryos are
shown. h, heart; ht, heart tube; oft, outflow tract. (D) Whole-mount in situ
hybridization to Hand1 transcripts at E10.5 in wild-type and
Hand1lacZ/loxP; MHC::Cre (mutant) embryos.
ba, branchial arch; lv, left ventricle; lm, lateral mesoderm. (E) Detection of
Hand1 and Hprt transcripts in RNA from the left ventricles
of wild-type and Hand1lacZ/loxP; MHC::Cre
(mutant) embryos at E9.5. Size markers are in the middle lane. Hand1
transcripts were undetectable in the mutant. Transcripts for Hprt
were detected as a control for RNA integrity and loading.
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