These Coll1a1-TetO-MSI2 (Coll-TetO-MSI2) double targeted mutation mice have doxycycline (tetracycline) inducible overexpression of human MSI2 (musashi homolog 2) and may be useful in conjunction with other oncogenic mutations for studies of the role of MSI2 in proliferation and differentiation of hematopoietic stem and myeloid progenitor cells in diseases such as human leukemia.
Rudolf Jaenisch, Whitehead Institute, Massachusetts Institute of Technology
Genetic Background | Generation |
---|---|
|
Allele Type | Gene Symbol | Gene Name |
---|---|---|
Targeted (Transactivator) | Gt(ROSA)26Sor | gene trap ROSA 26, Philippe Soriano |
Allele Type | Gene Symbol | Gene Name |
---|---|---|
Targeted (Inducible, Inserted expressed sequence, Humanized sequence, RMCE-ready) | Col1a1 | collagen, type I, alpha 1 |
These Coll1a1-TetO-MSI2 (Coll-TetO-MSI2) double mutant mice carry two targeted mutations: (1) an optimized form of reverse tetracycline controlled transactivator (rtTA*M2) in the Gt(ROSA)26Sor locus and (2) a tetracycline responsive element (tetO or TRE)-controlled human MSI2 (musashi homolog 2) gene in the Col1a1 locus. MSI2 plays a role in regulation of hematopoiesis. Aberrant MSI2 expression is associated with aggressive myeloid leukemia and poor prognosis in blast crisis chronic myelogenous leukemia. Doxycycline administration induces widespread overexpression of human MSI2 (musashi homolog 2), resulting in expansion of hematopoietic stem and progenitor cells.
While mutant mice treated continuously with doxycycline for 1 year exhibit increased spleen and liver weights, and a reduction in MSI2 induction over time, they do not develop leukemia. Mice that are homozygous for the targeted mutations and untreated with doxycycline are viable, fertile, normal in size and do not display any gross physical or behavioral abnormalities. Recipient control mice transplanted with bone marrow from these Coll1a1-TetO-MSI2 double mutant mice exhibit (with doxycycline treatment) increased hematopoietic stem and progenitor cells, increased hematocrit and mean corpuscular volume; and decreased myeloid progenitor, myeloid erythroid progenitor cells, neutrophils, lymphocytes and platelets.
The R26-M2rtTA targeted mutation has an optimized form of reverse tetracycline controlled transactivator (rtTA-M2) followed by a β-globin intron, polyA signal and PGK-puromycin cassette, all inserted downstream of the Gt(ROSA)26Sor promoter. The targeting vector was electroporated into (C57BL/6 x 129S4Sv/Jae)F1-derived V6.5 embryonic stem (ES) cells. Correctly targeted ES cells (called KH2) carrying the Gt(ROSA)26Sortm1(rtTA*M2)Jae allele, were retargeted to insert an FRT site-flanked PGK-Neo cassette and a promoterless hygromycin resistance cassette downstream of the Col1a1 locus. Correctly retargeted ES cells were selected and the FRT site-flanked PGK-Neo downstream of the Col1a1 locus was replaced by electroporating both a "flip-in plasmid" containing a PGK promoter, ATG start codon, FRT site, a splice acceptor-double polyA sequence, tetO (tetracycline responsive element) with a CMV minimal promoter, and SV40 polyA signal, upstream of the human MSI2 cDNA sequence, and a FLPe recombinase expressing vector to facilitate tetO-MSI2 recombination into the 3' UTR of the Col1a1 locus. The resulting correctly targeted ES cells, carrying rtTA*M2 in the Gt(ROSA)26Sor locus and tetO-MSI2 in the Col1a1 locus, were injected into recipient blastocysts. The mice were backcrossed to C57BL/6 for 5 generations.
Upon arrival at The Jackson Laboratory, the mice were crossed to C57BL/6J (Stock No. 000664) at least once to establish the colony.
Expressed Gene | rtTA, reverse tetracycline-controlled transactivator, E. coli |
---|---|
Site of Expression | Expresses an optimized rtTA protein (rtTA-M2). Inducible target gene expression is detected in liver, bone marrow, stomach, intestine, and skin, with lower levels in the heart, lungs, kidney, spleen, and thymus; no expression is detected in the brain and testes. |
Expressed Gene | MSI2, musashi RNA binding protein 2, human |
Site of Expression | MSI2 is normally expressed in hematopoietic stem cells. |
Allele Name | targeted mutation 1, Rudolf Jaenisch |
---|---|
Allele Type | Targeted (Transactivator) |
Allele Synonym(s) | Gt(ROSA)26Sortm1(M2rtTA)Jae; M2; M2-rtTA; R26-M2rtTA; R26-rtTA; Rosa26-M2rtTA |
Gene Symbol and Name | Gt(ROSA)26Sor, gene trap ROSA 26, Philippe Soriano |
Gene Synonym(s) | |
Expressed Gene | rtTA, reverse tetracycline-controlled transactivator, E. coli |
Site of Expression | Expresses an optimized rtTA protein (rtTA-M2). Inducible target gene expression is detected in liver, bone marrow, stomach, intestine, and skin, with lower levels in the heart, lungs, kidney, spleen, and thymus; no expression is detected in the brain and testes. |
Strain of Origin | (C57BL/6 x 129S4/SvJae)F1 |
Chromosome | 6 |
General Note | This mutation was created during the generation of mutant ES cell line KH2, which also contains a frt docking site downstream of the endogenous Col1a1 locus on Chr 11 (Col1a1 |
Molecular Note | An optimized form of reverse tetracycline controlled transactivator (rtTA-M2) was inserted downstream of the Gt(ROSA)26Sor promoter and was followed by a PGK-puro selection cassette. This mutant form of rtTA termed M2 has five amino acid substitutions in the tetR moiety of tTA: S12G, E19G, A56P, D148E and H179R. This mutated form of transactivator protein has increased doxycycline sensitivity. Mice have widespread expression of the rtTA-M2 protein. |
Mutations Made By | Rudolf Jaenisch, Whitehead Institute, Massachusetts Institute of Technology |
Allele Name | targeted mutation 6, Rudolf Jaenisch |
---|---|
Allele Type | Targeted (Inducible, Inserted expressed sequence, Humanized sequence, RMCE-ready) |
Allele Synonym(s) | Coll-TetO-MSI2 |
Gene Symbol and Name | Col1a1, collagen, type I, alpha 1 |
Gene Synonym(s) | |
Promoter | tetO, tet operator, |
Expressed Gene | MSI2, musashi RNA binding protein 2, human |
Site of Expression | MSI2 is normally expressed in hematopoietic stem cells. |
Strain of Origin | (C57BL/6 x 129S4/SvJae)F1 |
Chromosome | 11 |
General Note | The allele was made in KH2 cells that carry Gt(ROSA)26Sortm1(rtTA*M2)Jae and Col1a1tm2(tetO-Pou5f1)Jae (J:159351). |
Molecular Note | RMCE on KH2 cells inserted a cassette that contains a tetracycline operator and minimal CMV promoter drives inducible expression of the human cDNA preceded by a splice acceptor and polyadenylation sequence and followed by an FRT flanked PGK-neo cassette and promotorless hygro cassette into the 3'UTR. |
When maintaining a live colony, these mice can be bred as homozygotes for both mutant alleles.
When using the Coll-TetO-MSI2 mouse strain in a publication, please cite the originating article(s) and include JAX stock #014588 in your Materials and Methods section.
Facility Barrier Level Descriptions
Service/Product | Description | Price |
---|---|---|
Heterozygous for26Sor<tm1(rtTA*M2)Jae> , Heterozygous for Col1A1<tm6(tetO-MSI2)Jae> |
Frozen Mouse Embryo | B6.Cg-Gt(ROSA)26Sor<tm1(rtTA*M2)Jae> Col1a1<tm6(tetO-MSI2)Ja | $2595.00 |
Frozen Mouse Embryo | B6.Cg-Gt(ROSA)26Sor<tm1(rtTA*M2)Jae> Col1a1<tm6(tetO-MSI2)Ja | $2595.00 |
Frozen Mouse Embryo | B6.Cg-Gt(ROSA)26Sor<tm1(rtTA*M2)Jae> Col1a1<tm6(tetO-MSI2)Ja | $3373.50 |
Frozen Mouse Embryo | B6.Cg-Gt(ROSA)26Sor<tm1(rtTA*M2)Jae> Col1a1<tm6(tetO-MSI2)Ja | $3373.50 |
Terms are granted by individual review and stated on the customer invoice(s) and account statement. These transactions are payable in U.S. currency within the granted terms. Payment for services, products, shipping containers, and shipping costs that are rendered are expected within the payment terms indicated on the invoice or stated by contract. Invoices and account balances in arrears of stated terms may result in The Jackson Laboratory pursuing collection activities including but not limited to outside agencies and court filings.
The Jackson Laboratory has rigorous genetic quality control and mutant gene genotyping programs to ensure the genetic background of JAX® Mice strains as well as the genotypes of strains with identified molecular mutations. JAX® Mice strains are only made available to researchers after meeting our standards. However, the phenotype of each strain may not be fully characterized and/or captured in the strain data sheets. Therefore, we cannot guarantee a strain's phenotype will meet all expectations. To ensure that JAX® Mice will meet the needs of individual research projects or when requesting a strain that is new to your research, we suggest ordering and performing tests on a small number of mice to determine suitability for your particular project. We do not guarantee breeding performance and therefore suggest that investigators order more than one breeding pair to avoid delays in their research.
What information were you hoping to find through your search?
How easy was it to find what you were looking for?
We may wish to follow up with you. Enter your email if you are happy for us to connect and reachout to you with more questions.
Please Enter a Valid Email Address
Thank you for sharing your feedback! We are working on improving the JAX Mice search. Come back soon for exciting changes.