These Col1a1-tetO-Dnmt3b1::R26-M2rtTA mice may be used to regulate expression of Dnmt3b1 via a tetracycline-controlled transactivator protein system, and may be useful in studying the regulation of tumorigenesis.
Rudolf Jaenisch, Whitehead Institute, Massachusetts Institute of Technology
Genetic Background | Generation |
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Allele Type | Gene Symbol | Gene Name |
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Targeted (Transactivator) | Gt(ROSA)26Sor | gene trap ROSA 26, Philippe Soriano |
Allele Type | Gene Symbol | Gene Name |
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Targeted (Inducible, Inserted expressed sequence, RMCE-ready) | Col1a1 | collagen, type I, alpha 1 |
Mice homozygous for the Col1a1-tetO-Dnmt3b1::R26-M2rtTA targeted mutation are viable and fertile. Expression of the DNA methyltransferase 3B gene (Dnmt3b1) is controlled by the tet-responsive element (tetO). DNMT3B1 is a de novo methyltransferase which is able to methylate CpG sequences, a required step during development. Hypermethylation of DNA has been implicated in many neoplasms, altering gene expression patterns that contribute to neoplastic transformation, tumorigenesis, and tumor behavior. This strain also contains the optimized form of reverse tetracycline controlled transactivator (rtTA-M2). Expression of Dnmt3b1 can be regulated with the tetracycline analog doxycycline (dox) in these double mutant offspring.
When these mice are bred to C57BL/6J-ApcMin/J mice (see Stock No. 002020), DNMT3B1 expression in dox induced mice results an increase in intestinal tumor number and size as compared to ApcMin control mice.
A targeting vector was designed to insert frt-flanked PGK-Neo-pA 500 bp downstream of the 3' UTR of the Col1a1 locus. This construct was electroporated into (C57BL/6 x 129S4Sv/Jae)F1-derived V6.5 embryonic stem (ES) cells. Correctly targeted ES cells were identified. One of these ES cells clones, C10, was re-targeted to insert an optimized form of reverse tetracycline controlled transactivator (rtTA-M2) followed by a β-globin intron, polyA signal and PGK-puromycin cassette, all downstream of the Gt(ROSA)26Sor promoter. Correctly targeted ES cells were identified. One of these ES cell clones, KH2, was selected and the frt-flanked PGK-Neo-pA in the 3' UTR of the Col1a1 locus was replaced by co-injection of a tetO-Dnmt3b1 plasmid. The tetO-Dnmt3b1 plasmid contained a splice acceptor-double polyA sequence and the tetracycline responsive element (tetO) upstream of the DNA methyltransferase 3B gene.
The resulting ES cells, now targeted with rtTA-M2 in the Gt(ROSA)26Sor locus (Rosa26-rtTA) and tetO-Dnmt3b1 in the Col1a1 locus (Col1a1-tetO-Dnmt3b1), were injected into recipient blastocysts. The donating investigator stated that the resulting chimeric mice were bred together and were subsequently backcrossed to C57BL/6J mice (see SNP note below) for at least 7 generations to establish the double mutant colony.
Upon arrival at The Jackson Laboratory Repository, these mice were bred to C57BL/6J inbred mice (Stock No. 000664) for at least one generation.
A 32 SNP (single nucleotide polymorphism) panel analysis, with 27 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. While the 27 markers throughout the genome suggested a C57BL/6 genetic background, 4 of 5 markers that determine C57BL/6J from C57BL/6N were found to be segregating. These data suggest the mice sent to The Jackson Laboratory Repository were on a C57BL/6N genetic background.
Expressed Gene | rtTA, reverse tetracycline-controlled transactivator, E. coli |
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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 | Dnmt3b, DNA methyltransferase 3B, mouse, laboratory |
Site of Expression |
Allele Name | targeted mutation 1, Rudolf Jaenisch |
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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 9, Rudolf Jaenisch |
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Allele Type | Targeted (Inducible, Inserted expressed sequence, RMCE-ready) |
Allele Synonym(s) | tetO-Dnmt3b1 |
Gene Symbol and Name | Col1a1, collagen, type I, alpha 1 |
Gene Synonym(s) | |
Promoter | tetO, tet operator, |
Expressed Gene | Dnmt3b, DNA methyltransferase 3B, mouse, laboratory |
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 splice acceptor-double polyA sequence and the tetracycline responsive element (tetO) upstream of the DNA methyltransferase 3B gene isoform 1 ( |
When maintaining a live colony, homozygous mice may be bred together.
When using the Col1a1-tetO-Dnmt3b1::R26-M2rtTA mouse strain in a publication, please cite the originating article(s) and include JAX stock #017983 in your Materials and Methods section.
Facility Barrier Level Descriptions
Service/Product | Description | Price |
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Heterozygous for Col1a1<tm9(tetO-Dnmt3b1_i1)Jae> |
Frozen Mouse Embryo | B6.Cg-Gt(ROSA)26Sor<tm1(rtTA*M2)Jae> Col1a1<tm9(tetO-Dnmt3b_ | $2595.00 |
Frozen Mouse Embryo | B6.Cg-Gt(ROSA)26Sor<tm1(rtTA*M2)Jae> Col1a1<tm9(tetO-Dnmt3b_ | $2595.00 |
Frozen Mouse Embryo | B6.Cg-Gt(ROSA)26Sor<tm1(rtTA*M2)Jae> Col1a1<tm9(tetO-Dnmt3b_ | $3373.50 |
Frozen Mouse Embryo | B6.Cg-Gt(ROSA)26Sor<tm1(rtTA*M2)Jae> Col1a1<tm9(tetO-Dnmt3b_ | $3373.50 |
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