C57BL/6J-Gt(ROSA)26Sor^{tm1(CAG-RUNX1*,-EGFP)Dez}/J

Stock number: 041734
Product name: Rosa26-AE9a knock-in
Research areas: Cancer Research, Cell Biology Research, Neurobiology Research, Research Tools

Description

These Rosa26-AE9a knock-in (KI) mice express a Cre-inducible human RUNX1 AE9a mutant allele, as well as an EGFP, both driven by a CAG promoter from the Gt(ROSA)26Sor locus. These mice may be useful for studying AML and other blood cancers.

Detailed description

Runt related transcription factor 1 (RUNX1) is crucial for hematopoietic cell development. RUNX1 also controls stem cell fate and proliferation in various organs and is involved in development of blood vessels, muscles, neurons, and epithelial tissues. Runx1 expression is regulated by two promoters and alternative splicing results in multiple isoforms. In humans, RUNX1 is a frequent target of translocations or mutations implicated in leukemias and myelodysplastic syndrome (MDS). The most common of these, t(8;21), occurs at a chromosomal translocation hot spot in the human RUNX1 locus resulting in expression of the RUNX1-RUNX1T1 (RUNX1 translocation partner 1) fusion oncoprotein, also referred to as AML1-ETO (AE), which lacks all C-terminal functional domains. Overexpression of a C-terminal truncated variant of the fusion protein, AML1-ETO9a (AE9a) is also detected in acute myeloid leukemia (AML) patients. These Rosa26-AE9a knock-in (KI) mice co-express a Cre-inducible human RUNX1 AE9a mutant allele, as well as an enhanced green fluorescent protein (EGFP), both driven by a CAG promoter from the Gt(ROSA)26Sor locus.

Heterozygous mice are viable and fertile with no overt phenotype. The donating laboratory has not attempted to generate homozygous mice. When bred to mice that express tissue-specific Cre recombinase, resulting offspring will express the human RUNX1 AE9a mutant allele in cre-expressing tissues. EGFP fluorescence is detectable by flow cytometry and immunostaining. The donating laboratory bred these mice to a Vav-iCre (Stock No. 008610) mouse line and confirmed AE9a expression solely in the bone marrow, spleen, and blood. Rosa26-AE9a KI mice do develop AML by ~8-9 months of age. These mice may be useful for studying AML and other blood cancers.

Development

The Rosa26-AE9a knock-in (KI) allele was designed to introduce a CMV-IE enhancer/chicken beta-actin/rabbit beta-globin hybrid promoter (CAG), a loxP-flanked STOP (PGK-Neo-PGKpA) cassette, an HA-tagged human runt related transcription factor 1 (RUNX1) sequence, harboring the AE9a mutation, followed by an FRT-flanked internal ribosome entry site (IRES) and enhanced green fluorescent protein (EGFP) cassette, and polyA signal, all inserted into the Gt(ROSA)26Sor locus on Chromosome 6. The construct was electroporated into C57BL/6J-derived embryonic stem (ES) cells and correctly targeted ES cells were injected into recipient blastocysts. Resulting Rosa26-AE9a mice were maintained on a C57BL/6J background by the donating laboratory. To establish our live colony, an aliquot of frozen sperm was used to fertilize C57BL/6J (Stock No. 000664) oocytes.

Allele

Symbol: Gt(ROSA)26Sor^{tm1(CAG-RUNX1*,-EGFP)Dez}
Name: targeted mutation 1, Dong-Er Zhang
MGI accession ID: MGI:8353621
Generation method: Targeted
Attributes: Conditional ready (e.g. floxed); Reporter; Inserted expressed sequence; Epitope tag
Synonyms: Rosa26-AE9a
Marker symbol: Gt(ROSA)26Sor
Marker name: gene trap ROSA 26, Philippe Soriano
Marker synonyms: beta geo; Gtrgeo26; Gtrosa26; R26; ROSA26; SETD5-AS1; Thumpd3as1
Chromosome: 6
Strain of origin: C57BL/6J
Molecular note: The allele was designed to introduce a CMV-IE enhancer/chicken beta-actin/rabbit beta-globin hybrid promoter (CAG), a loxP-flanked STOP (PGK-Neo-PGKpA) cassette, an HA-tagged human runt related transcription factor 1 (RUNX1) sequence, harboring the AE9a mutation, followed by an FRT-flanked internal ribosome entry site (IRES) and enhanced green fluorescent protein (EGFP) cassette, and polyA signal, all inserted into the Gt(ROSA)26Sor locus on Chromosome 6.