129S-H3f3a^tm1.1Mnn/J

Stock number: 021788
Product name: H3f3a^c , H3f3a floxed (cko+YFP)
Research areas: Cell Biology Research, Research Tools

Description

These H3f3a^c/c mutant mice possess loxP sites flanking a wildtype H3 histone isoform 3 (H3.3) coding sequence and STOP sequence. When bred to mice expressing cre-recombinase, removal of the floxed sequence results in YFP expression. This strain may be useful for studying the role of histone H3.3 during development.

Detailed description

The H3f3a^c/c targeting vector contains, from 5' to 3', a synthetic splice acceptor, a loxP-site, a wildtype H3 histone isoform 3 (H3.3) coding sequence, H3f3a (H3 histone, family 3A) 3' UTR, and an SV40 polyadenylation (polyA) sequence. This was followed by a second loxP site, an ATG start codon, the venus variant of yellow fluorescent protein (YFP), and another SV40 polyA sequence. This vector replaced part of exon 2 of the H3f3a gene. H3f3a encodes the histone H3 isoform, H3.3, involved in the regulation of chromatin activity during DNA replication. Homozygous H3f3a^c/c mice are viable and fertile, and express wildtype H3f3a. In the absence of Cre, YFP expression is prevented by the H3.3 coding sequence. When bred to mice that express Cre recombinase, resulting offspring will have the wildtype loxP-flanked H3.3 coding sequence deleted in the cre-expressing tissues, allowing YFP to be expressed.

Development

A targeting vector was designed to contain, from 5' to 3', a synthetic splice acceptor, a loxP site, an ATG start codon, a wildtype H3 histone isoform 3 (H3.3) coding sequence, H3f3a (H3 histone, family 3A) 3' UTR, and an SV40 polyadenylation (polyA) sequence. This was followed by a downstream frt-flanked neomycin resistance (neo) cassette, and a second loxP site, an ATG start codon, the venus variant of yellow fluorescent protein (YFP), and another SV40 polyA sequence. This vector replaced part of exon 2 of the H3f3a gene. The construct was electroporated into 129S1/SvImJ-derived A2 embryonic stem (ES) cells. Correctly targeted ES cells were injected into (C57BL/6 J x CBA/CaH) blastocysts and resulting chimeric mice were bred with 129S1;129S4-Gt(ROSA)26Sor^tm1(FLP1)Dym/J transgenic mice to delete the neo cassette. Resulting offspring were crossed to remove the Flp-expressing transgene, resulting in a colony homozygous for the conditional-H3f3a (H3f3a^c/c) allele. These mice were bred to 129S1/SvImJ mice (Stock No. 002448) for at least 5 generations. Upon arrival, mice were bred to 129S1/SvImJ inbred mice for at least one generation to establish the colony.

Allele

Symbol: H3f3a^tm1.1Mnn
Name: targeted mutation 1.1, Jeff Mann
MGI accession ID: MGI:5474375
Generation method: Targeted
Attributes: Conditional ready (e.g. floxed); No functional change
Synonyms: H3f3a^c
Marker symbol: H3f3a
Marker name: H3 histone, family 3A
Marker synonyms: BRYLIB1; BRYLIB2; H3.3A; H3.3B; H3-3a; H3-3B; H3F3; H3F3A; H3F3B
Chromosome: 1
Strain of origin: 129S1/SvImJ
Molecular note: A targeting vector was designed to contain, from 5' to 3', a synthetic splice acceptor, a loxP site, an ATG start codon, a wild-type H3 histone isoform 3 (H3.3) coding sequence, H3f3a (H3 histone, family 3A) 3' UTR, and an SV40 polyadenylation (polyA) sequence. This was followed by a downstream frt-flanked neomycin resistance (neo) cassette, and a second loxP site, an ATG start codon, the venus variant of yellow fluorescent protein (YFP), and another SV40 polyA sequence. This vector replaced part of exon 2. Flp-mediated recombination removed the neo cassette.