B6;129-Ncoa1^tm1.1So/J

Stock number: 035498
Product name: SRC-1^P1278S
Research areas: Cancer Research, Cell Biology Research, Research Tools

Description

SRC-1^{P1278S/P1278S} knock-in/floxed mice carry a proline to serine mutation (P1278S) in exon 16 of the nuclear receptor coactivator 1 (Ncoa1) gene, similar to a mutation found in a small cohort of bone metastases from breast cancer patients. Also, exons 15-16 are floxed.

Detailed description

SRC-1^{P1278S/P1278S} knock-in/floxed mice carry a proline to serine mutation (P1278S) in exon 16 of the nuclear receptor coactivator 1 (Ncoa1) gene, similar to a mutation found in a small cohort of bone metastases from breast cancer patients. Exon 16 encodes the activation domain 2 of SRC-1. Also, exons 15-16 are floxed. Mice homozygous for this knock-in SRC-1 allele exhibit an increase in tamoxifen-induced bone protection after ovariectomy.

Homozygous mice are viable and fertile. When bred to mice that express tissue-specific Cre recombinase, resulting offspring will have exons 15-16 of SRC-1 deleted in the cre-expressing tissues.

Development

The SRC-1^P1278S targeting construct was designed to insert a loxP site downstream of exon 14 of the nuclear receptor coactivator 1 (Ncoa1) gene, and an FRT-flanked neomycin (neo) resistance cassette, followed by a second loxP site downstream of exon 16. A CCA to TCA mutation was introduced in exon 16, encoding activation domain 2, converting a proline to a serine (P1278S) similar to a mutation found in a small cohort of bone metastases from breast cancer patients. This targeting construct was electroporated into 129-derived embryonic stem (ES) cells and correctly targeted ES cells were injected into albino C57BL/6 blastocysts. Chimeric mice were bred together and subsequently mice were bred to ROSA26::FLPe transgenic mice (Stock No. 009086) to remove the neo cassette. The resulting SRC-1^{P1278S/P1278S} offspring were backcrossed to C57BL/6J mice for at least 5 generations by the donating lab (see SNP note below). Upon arrival at The Jackson Laboratory, sperm was cryopreserved. To establish our live colony, an aliquot of frozen sperm was used to fertilize C57BL/6J oocytes (Stock No. 000664).

In 2020, 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. Five of the 43 markers throughout the genome were segregating with 129, suggesting an incomplete backcross.

Allele

Symbol: Ncoa1^tm1.1So
Name: targeted mutation 1.1, Steffi Oesterreich
MGI accession ID: MGI:6442000
Generation method: Targeted
Attributes: Conditional ready (e.g. floxed); Humanized sequence
Synonyms: NCOA1 P1278S SNP; SRC1^P1278S
Marker symbol: Ncoa1
Marker name: nuclear receptor coactivator 1
Marker synonyms: bHLHe42; bHLHe74; F-SRC-1; KAT13A; RIP160; SRC1; SRC-1; SRC-a/NCoA-1; steroid receptor coactivator-1
Chromosome: 12
Strain of origin: 129
Molecular note: The targeting construct is designed to insert a loxP site downstream of exon 14 and an FRT-flanked neomycin (neo) resistance cassette, followed by a second loxP site downstream of exon 16. A CCA to TCA mutation was introduced in exon 16, encoding activation domain 2, converting a proline to a serine (P1278S) similar to a mutation found in a small cohort of bone metastases from breast cancer patients. Flp-mediated recombination removed the FRT-flanked neomycin cassette.