STOCK Tg(LCR-HBG2*/HBG1)100Hhb/J

Stock number: 031709
Product name: high gamma, γH
Research areas: Hematological Research, Research Tools

Description

These high gamma transgenic mice carry the γH transgene and may be suitable for use in studies related to sickle cell anemia and hemoglobinopathy.

Detailed description

These transgenic mice carry the high gamma, γH, transgene. The originating investigator reports there are multiple copies of the transgene. High gamma transgenic mice exhibit decreased oxygen affinity (p50 of 24.9 mm Hg) compared to wildtype C57BL/6J controls (p50 of 40mm Hg), and have decreased levels of 2,3 diphosphoglycerol in red blood cells. Hemizygotes are viable and fertile. Homozygous viability/fertility has not been tested (February 2018). The Donating Investigator suggests extending weaning for smaller pups.

When crossed to Berkeley model mice, B6.Cg-Hba^tm1Paz Hbb^tm1Tow Tg(HBA-HBBs)41Paz (Stock No. 031405), the resulting adult mice exhibit smaller spleens and longer lifespans than Berkeley model mice. Genotyping assays developed by The Jackson Laboratory were validated by HPLC analysis.

Development

The high gamma, γH, transgene contains a PCR-generated fragment of HS4 of the LCR of the human HBB gene (951-2204) upstream of a mutant human HBG1 (^Gγ globin) gene with a T at position -158 relative to the cap site, inserted into a 12.4-kb fragment of the human HBB gene (32837-45203) including the 3' enhancer and both ^Gγ globin and ^Aγ globin genes. The high gamma transgene was microinjected into unspecified fertilized oocytes. Founder line G100 was subsequently established and found to have multiple copies of the transgene. The mice were backcrossed to C57BL/6J mice for more than 6 generations. The mice were then crossed to B6.Cg-Hba^tm1Paz Hbb^tm1Tow Tg(HBA-HBBs)41Paz/J, Berkeley model mice on the congenic C57BL/6J background. Upon arrival, sperm was cryopreserved. To establish our live colony, an aliquot of frozen sperm was used to fertilize C57BL/6J oocytes (Stock No. 000664), and bred to remove the Hba^tm1Paz Hbb^tm1Tow alleles and Tg(HBA-HBBs)41Paz transgene.

In 2018, 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 rederived living colony at The Jackson Laboratory Repository. The SNP analysis revealed that this strain is on a mixed STOCK background.

Allele

Symbol: Tg(LCR-HBG2*/HBG1)100Hhb
Name: transgene insertion 100, Henny Billett
MGI accession ID: MGI:6277080
Generation method: Transgenic
Attributes: Inserted expressed sequence; Humanized sequence
Synonyms: gammaH
Marker symbol: Tg(LCR-HBG2*/HBG1)100Hhb
Marker name: transgene insertion 100, Henny Billett
Chromosome: UN
Strain of origin: Not Specified
Molecular note: The transgene contains a PCR-generated fragment of HS4 of the locus control region (LCR) of the human HBB gene region (951-2204) inserted into a 12.4-kb fragment of the human HBB gene region (32837-45203) including the mutant human HBG2 gene with a T at position -158 relative to the cap site, the HBG1 gene and the 3' enhancer. Founder line G100 was established.