STOCK Tg(LCR-HBG2*/HBG1)203Hhb/J

Stock number: 031708
Product name: medium gamma, γM
Research areas: Hematological Research, Research Tools

Description

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

Detailed description

These transgenic mice carry the medium gamma, γM, transgene. The originating investigator reports there are multiple copies of the transgene. 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 a less severe phenotype: postnatal expression of γ-globin (HbF) increases with time to 21% at 9 weeks of age compared to Berkeley mice, B6.Cg-Hba^tm1Paz Hbb^tm1Tow Tg(HBA-HBBs)41Paz/J, (<1%); and hematocrit count increases to 41, compared to 29 observed in Berkeley mice. Genotyping assays developed by The Jackson Laboratory were validated by HPLC analysis.

Development

The medium gamma, γM, transgene, designed by Drs. Bernard G. Forget (Yale University School of Medicine) and John Gilman (Albert Einstein College of Medicine), contains a PCR-generated 4.3kb fragment of the human HBB gene (912-5200) containing HS3 and HS4 of the LCR 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 γM transgene was microinjected into unspecified fertilized oocytes. Founder line G203 was subsequently established and found to have multiple copies of the transgene. The medium gamma transgenic mice were then bred to the Berkeley model mice on a congenic C57BL/6J background: B6.Cg-Hba^tm1PazHbb^tm1Tow Tg(HBA-HBBs)41Paz. 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^tm1PazHbb^tm1Tow alleles and Tg(HBA-HBBs)41Paz transgene.

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 rederived living colony at The Jackson Laboratory Repository. The results suggest the mice sent to The Jackson Laboratory Repository were on a mixed C57BL/6 and unknown genetic background.

Allele

Symbol: Tg(LCR-HBG2*/HBG1)203Hhb
Name: transgene insertion 203, Henny Billett
MGI accession ID: MGI:6276606
Generation method: Transgenic
Attributes: Inserted expressed sequence; Humanized sequence
Synonyms: gammaM
Marker symbol: Tg(LCR-HBG2*/HBG1)203Hhb
Marker name: transgene insertion 203, Henny Billett
Chromosome: UN
Strain of origin: Not Specified
Molecular note: The transgene contains a PCR-generated 4.3kb fragment of the human HBB gene (912-5200) containing HS3 and HS4 of the human beta-globin locus control regions (LCR) inserted into a 12.4-kb fragment of the human HBB gene (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 G203 was established and found to have multiple copies of the transgene.