B6.Cg-Gas1^{em1(icre/ERT2)Nono}/J

Stock number: 038342
Product name: Gas1-creER
Research areas: Apoptosis Research, Cancer Research, Cell Biology Research, Research Tools

Description

Gas1-creER knock-in mice express an iCreER fusion protein driven by the growth arrest specific 1 (Gas1) endogenous promoter. When these mice are bred with mice containing a loxP-flanked sequence of interest, tamoxifen-inducible, Cre-mediated recombination will result in deletion of the floxed sequences in the Gas1-expressing cells of the offspring. These mice may be useful for studying cellular proliferation, apoptosis, embryonic development and tumor suppression in certain cancers.

Detailed description

Gas1-creER knock-in mice express a tamoxifen-inducible, codon-improved, iCre recombinase (iCre/ERT2) under the control of the growth arrest specific 1 (Gas1) endogenous promoter, while retaining endogenous Gas1 function. Growth arrest-specific 1 (GAS1) is an abundant cell membrane protein that plays a role in growth suppression.

Restricted to the cytoplasm, iCre-ERT can only gain access to the nuclear compartment after exposure to tamoxifen. When Gas1-creER mice are bred with mice containing loxP-flanked sequences, tamoxifen-inducible Cre-mediated recombination will result in deletion of the floxed sequences in the cre-expressing cells of the offspring. The donating investigator indicates that while their Gas1-creER colony is maintained by crossing heterozygous mice to C57BL/6 (wildtype) mice, homozygous mice are also viable and fertile with no observable phenotype. The donating laboratory crossed the Gas1-creER mice to an Ai14 R26-tdTomato reporter mouse (Stock No. 007914) and confirmed expression, after tamoxifen induction, of the fluorescence reporter in non-hematopoietic cells of the femur. The tdTomato reporter was noted as being highly expressed in a novel cell type named osteoblast-reticular transitional (ORT) cells and localized to the endosteal space. These Gas1-creER knock-in mice may be useful for studying the role of Gas1 in inhibition of cellular proliferation, apoptosis and tumor growth in certain cancers.

Development

The Gas1-creER knock-in allele was generated using CRISPR/Cas9 endonuclease-mediated genome editing. A guide RNA (5'-CCTGGAGCAGCCTGCAGCAG-3' (PAM = GGG)) was used to introduce P2A-iCreERT2-bGH polyA sequences into the 3' UTR region of the growth arrest specific 1 (Gas1) gene on chromosome 13. Gas1 transcript Gas1-201 (ENSMUST00000065086.6) was used as reference for the exon number and the guide sequences. The sgRNAs, ssDNA and the Cas9 nuclease mRNA were introduced into the cytoplasm of zygotes derived from B6SJLF1/J (Stock No. 100012) mice with well recognized pronuclei. Correctly targeted pups were identified by PCR and sequencing. The resulting Gas1-creER knock-in mice were bred to C57BL/6 mice for at least 11 generations. To establish our live colony, an aliquot of frozen sperm was used to fertilize C57BL/6J (Stock No. 000664) oocytes.

Allele

Symbol: Gas1^{em1(icre/ERT2)Nono}
Name: endonuclease-mediated mutation 1, Noriaki Ono
MGI accession ID: MGI:7484373
Generation method: Endonuclease-mediated
Attributes: Recombinase-expressing; Inducible
Synonyms: Gas1-creER
Marker symbol: Gas1
Marker name: growth arrest specific 1
Marker synonyms: Gas-1
Chromosome: 13
Strain of origin: (C57BL/6J x SJL/J)F1
Molecular note: using CRISPR/Cas9 endonuclease-mediated genome editing, P2A-iCreERT2-bGH polyA sequences were inserted into the 3' UTR region of the growth arrest specific 1 (Gas1) gene on chromosome 13.