STOCK Hcn4^tm3Sev/J

Stock number: 024284
Product name: HCN4nLacZ/H2BGFP
Research areas: Cardiovascular Research, Research Tools

Description

HCN4nLacZ/H2BGFP mice are useful for visualizing cells in the first heart field of the developing cardiac crescent.

Detailed description

Heterozygous mice are viable and fertile, homozygotes are embryonic lethal. The HCN4nLacZ/H2BGFP allele expresses a floxed nuclear lacZ and a H2BGFP fusion protein from the hyperpolarization-activated, cyclic nucleotide-gated K+ 4 (HCN4) promoter/enhancer elements. The frt-flanked neo is still present in this strain. HCN4 is expressed in the first heart field of differentiating myocytes in the developing cardiac crescent. These mice express in the first heart field of the cardiac conduction system (CCS). When these mice are bred to mice that express Cre recombinase, resulting offspring will have the floxed-lacZ sequence removed, resulting in EGFP fluorescence in the CCS.

Development

A targeting vector was designed to insert a floxed nuclear lacZ followed by H2BGFP fusion protein, and frt-flanked neomycin resistance selection cassette, immediately before endogenous start codon of hyperpolarization-activated, cyclic nucleotide-gated K+ 4 (HCN4) gene. Specifically, the H2BGFP fusion protein consists of the coding sequence for the human HIST1H2BB gene [histone 1 H2bb] followed C-terminally by Enhanced Green Fluorescent Protein gene. This construct was electroporated into embryonic stem (ES) cells. Correctly targeted ES cells were injected into recipient blastocysts and chimeric males were bred. These mice were maintained on a Black Swiss outbred background. Upon arrival at The Jackson Laboratory, mice were bred with C57BL/6J inbred mice (Stock No. 000664) for at least one generation to establish the colony.

Allele

Symbol: Hcn4^tm3Sev
Name: targeted mutation 3, Sylvia Evans
MGI accession ID: MGI:5529677
Generation method: Targeted
Attributes: Reporter; Null/Knockout; Humanized sequence
Marker symbol: Hcn4
Marker name: hyperpolarization-activated, cyclic nucleotide-gated K+ 4
Chromosome: 9
Strain of origin: Not Specified
Expressed genes: lacZ,beta-galactosidase,E. coli; GFP,Green Fluorescent Protein,
Site of expression: Mice carrying this allele express a floxed nuclear lacZ and a H2BGFP fusion protein in the first heart field of the cardiac conduction system.
Molecular note: A targeting vector was designed to insert a floxed nuclear lacZ followed by H2BGFP fusion protein, and frt-flanked neomycin resistance selection cassette, immediately before the endogenous start codon. The H2BGFP fusion protein consists of the coding sequence for the human HIST1H2BB gene [histone 1 H2bb] followed C-terminally by Enhanced Green Fluorescent Protein (EGFP) gene.