B6.Cg-Tg(Dct-rtTA2S*M2)B8Mrln Tg(tetO-HIST1H2BJ/GFP)47Efu/J

Stock number: 038300
Product name: iDct-GFP , Dct-rtTA/TRE-H2B-GFP
Research areas: Research Tools

Description

iDct-GFP mice target tetracycline/doxycycline-inducible green fluorescent protein (GFP) expression to melanocytes during all embryonic, neonatal, and adult developmental stages. This enables the sorting, tracking, and study of melanocytes, their associated diseases, and the development of melanoma when crossing with strains carrying melanomagenesis alleles.

Detailed description

iDct-GFP mice target tetracycline/doxycycline-inducible green fluorescent protein (GFP) expression to melanocyte nuclei during all embryonic, neonatal, and adult developmental stages. This enables the sorting, tracking, and study of melanocytes, their associated diseases, and the development of melanoma when crossing with strains carrying melanomagenesis alleles.

The iDct-GFP strain was created through crosses of two independently created transgenic strains of mice (PMID: 21248750, 21392368). The Dct-rtTA-tg allele expresses reverse tetracycline-controlled transactivator (rtTA2s-M2) from the mouse dopachrome tautomerase (Dct) promoter (PubMed: 21248750). RtTA2s-M2 functions at a much lower concentration of doxycycline than the original form of rtTA, is more stable in eukaryotic cells, and does not cause leaky background expression without doxycycline. The TRE-H2BGFP-tg allele (also found in Stock No. 005104) expresses a human histone 1, H2BJ-GFP fusion protein from the tetracycline response element (TRE; tetO) (PubMed 14671312). Administration of doxycycline to compound mutants incorporating these two alleles activates the expression of GFP in cells expressing Dct, an enzyme involved with melanin biosynthesis.

Doxycycline-induced iDct-GFP adult mice show expression of GFP in cells located in the bulb, the bulge, and the outer root sheath areas of the hair follicles, where melanocytes reside. Immunostaining has confirmed that GFP is exclusively expressed in DCT positive cells. No leaky background expression in the absence of doxycycline has been noted. GFP expression responds quickly, within 12–18 hours, to a single intraperitoneal injection of doxycycline.

It should be noted that TRE-H2BGFP transgenic mice have anomalous expression of H2BGFP in the hematopoietic stem cells of the bone marrow. Although this is not seen in the skin, appropriate controls should be performed when analyzing H2BGFP expression in other anatomic locations.

See Development section for note on the Cdh23 locus (wildtype vs. age related hearing loss allele) of cryorecovered mice.

Development

The iDct-GFP strain was created from the intercross (PubMed: 21248750 and 21392368) of two independently created transgenic alleles: Dct-rtTA-tg and TRE-H2BGFP. The compound mutant was backcrossed to C57BL/6N for more than 10 generations by the donating laboratory.

In brief:
Dct-rtTA-tg: The mouse Dct (dopachrome tautomerase) promoter drives expression of the reverse tetracycline-activated transactivator rtTA2s-M2 fragment and human beta globin 3'UTR. This mutation was originally created in FVB embryos. This is founder line number B8. (PubMed: 21248750)

TRE-H2BGFP: A fusion of human histone 1, H2bj protein and green fluorescent protein (HIST1H2BJ/GFP) is under the control of a tetracycline-responsive element (TRE) and cytomegalovirus regulatory elements (mCMV). This mutation was originally created in CD1 embryos. (PubMed 14671312)

Upon arrival at The Jackson Laboratory in 2023, sperm was cryopreserved. Cryorecovery/live colony development will involve C57BL/6NJ (Stock No. 005304).

In 2023, a 48 SNP (single nucleotide polymorphism) panel analysis, with 43 markers covering all 19 chromosomes and the X chromosome, as well as 5 markers that distinguish between the C57BL/6J and C57BL/6N substrains, was performed on the males sent to The Jackson Laboratory Repository (and used to cryopreserve sperm). All the markers were C57BL/6N allele-type with the exception of the Chromosome 10 SNP in the Cdh23 locus that was homozygous for the functional allele Cdh23⁺ rather than the age related hearing loss allele Cdh23^ahl normally present for C57BL/6. Cryorecovery from this frozen sperm using C57BL/6NJ oocytes will result in mice heterozygous at that locus (Cdh23^ahl/+).

Allele

Symbol: Tg(tetO-HIST1H2BJ/GFP)47Efu
Name: transgene insertion 47, Elaine Fuchs
MGI accession ID: MGI:3044190
Generation method: Transgenic
Attributes: Inducible; Inserted expressed sequence
Synonyms: pTRE-H2BGFP; tetO-H2bGFP; TetO-H2B-GFP; TRE-EGFP; TRE-H2B-GFP; TRE-mCMV- H2B-GFP
Marker symbol: Tg(tetO-HIST1H2BJ/GFP)47Efu
Marker name: transgene insertion 47, Elaine Fuchs
Marker synonyms: pTRE-H2BGFP; tetO-H2bGFP
Chromosome: UN
Strain of origin: CD-1
Expressed genes: H2BC11,H2B clustered histone 11,human; GFP,Green Fluorescent Protein,
Site of expression: These mice express the human histone 1, H2BJ, protein (HIST1H2BJ) and Green Fluorescent Protein (GFP) fusion protein, HIST1H2BJ/GFP, under the control of a tetracycline-responsive promoter element (TRE;tetO). When crossed with mice expressing a tetracycline-transactivator protein (rtTA) or tetracycline-controlled transactivator protein (tTA), tissue specific transgene expression in the bitransgenic progeny can be regulated by the administration of the tetracycline analog, doxycycline. Potential constitutive transgene expression should be examined in tissues of interest.
Molecular note: This transgene contains a sequence encoding the human histone 1, H2bj, protein (HIST1H2BJ), and green fluorescent protein fusion protein, HIST1H2BJ/GFP, under the control of a tetracycline-responsive promoter element (TRE) and cytomegalovirus regulatory elements (mCMV).

Allele

Symbol: Tg(Dct-rtTA2S*M2)B8Mrln
Name: transgene insertion B8, Glenn Merlino
MGI accession ID: MGI:5784700
Generation method: Transgenic
Attributes: Transactivator
Synonyms: Dct-rtTA
Marker symbol: Tg(Dct-rtTA2S*M2)B8Mrln
Marker name: transgene insertion, Glenn Merlino
Chromosome: UN
Strain of origin: FVB/N
Molecular note: The mouse Dct promoter drives expression of the reverse tetracycline-activated transactivator rtTA2s-M2 fragment. No lines were specified.