These humanized hCYP1A1_1A2_Cyp1a2/Cyp1a1(-/-) mice may useful in drug or carcinogen metabolism research; specifically as a model for human risk assessment studies involving drug or environmental toxicants that may be substrates for cytochrome P450 family members.
Daniel W Nebert, University of Cincinnati Medical Center
Genetic Background | Generation |
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Allele Type | Gene Symbol | Gene Name |
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Targeted (Null/Knockout) | Del(9Cyp1a2-Cyp1a1)1Dwn | deletion, Chr 9, Daniel W Nebert 1 |
Allele Type |
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Transgenic (Inserted expressed sequence, Humanized sequence) |
Mice homozygous for the Cyp1a2/Cyp1a1(-) targeted allele and carrying the hCYP1A1_1A2 transgene are viable and fertile with normal lifespan. As the Cyp1a2/Cyp1a1(-) targeted allele lacks the coding regions of both Cyp1a1 and Cyp1a2 genes, no expression of either gene transcript or protein is observed in liver or small intestine by quantitative RT-PCR or Western blot, respectively. Transgene expression of the orthologous human genes is observed in the same tissues. While oral benzo[alpha]pyrene (BaP) treatment of Cyp1a2/Cyp1a1(-/-) mutant mice leads to BaP-induced immunosuppression and sickness, the presence of the human CYP1A orthologs in these mice minimizes/prevents such toxicity. These humanized hCYP1A1_1A2_Cyp1a2/Cyp1a1(-/-) mice may useful in drug or carcinogen metabolism research; specifically as a model for human risk assessment studies involving drug or environmental toxicants that may be substrates for cytochrome P450 family members.
"Humanized" hCYP1A1_1A2_Cyp1a1/1a2(-/-) mice have interchromosal deletion of a segment of mouse chromosome 9 including the majority of the closely-linked Cyp1a2 and Cyp1a1 genes and also harbor a human CYP1A2 and CYP1A1transgene. Generation of these mice required multiple combinations of other targeting strategies. In detail, the Cyp1a2(t) targeted allele was first generated using a targeting vector designed to insert a loxP-flanked PGK-NEO cassette 350 bp downstream of the endogenous stop codon (about 60 bp 3' of exon 7). Following electroporation into 129S6/SvEvTac-derived embryonic stem (ES) cells and ES cell microinjection into C57BL/6 blastocysts, the chimera's were bred with C57BL/6 to generate Cyp1a2(t) mutant mice. To create the Cyp1a1 targeted allele, a targeting vector was designed to insert a loxP-flanked hypoxanthine phosphoribosyltransferase (HPRT) minigene in intron 1 and a loxP site downstream of the termination codon in exon 7. Following electroporation into 129P2/OlaHsd-derived E14TG2a (HPRT-) ES cells, ES cells were microinjected into the blastocoele cavity of C57BL/6J embryos, and chimeric males were bred with C57BL/6J females. These Cyp1a1(t) mutant mice were then bred to a Cre-recombinase strain (CAGGS-CRE, mixed C57BL/6J and FVB/J genetic background). Transgenic offspring found to be heterozygous for the floxed null Cyp1a1(-) allele (containing only exon 1, a portion of intron 1, and one remaining loxP site in the 3' UTR) and hemizygous for CAGGS-CRE were bred to mice heterozygous for the Cyp1a2(t) allele. Mutant mice (Cyp1a2(t), Cyp1a1(-), CAGGS-CRE) were bred to C57BL/6. Because of the close genomic position of these two genes, offspring could be selected which had undergone Cre recombinase-mediated interchromosomal recombination between the loxP sites 3' beyond the stop codons of the Cyp1a2 and Cyp1a1 genes. Such mice were backcrossed to C57BL/6 for 10 generations (while selecting against the Cre transgene) to generate Cyp1a1/1a2 mutant mice. To create "humanized" CYP1A1 and CYP1A2 transgenic mice (hCYP1A1_1A2), a single copy of the 180-kb human CYP1A1_CYP1A2 locus-containing BAC-H (BAC Human CTB clone 31H21: including the 23.3 kb spacer region, 90 kb of CYP1A2 3'-flanking region and 53 kb of CYP1A1 3' flanking region) was microinjected into (C57BL/6JxDBA/2J)F1 oocytes. Founder mice having a single copy of BAC-H were identified and then backcrossed for 10 generations to C57BL/6 mice. These hCYP1A1_1A2 transgenic mice were next bred with Cyp1a1/1a2 mutant mice, generating the final mutant strain; "humanized" hCYP1A1_1A2_Cyp1a1/1a2(-/-) mice.
NOTE: A 22 SNP (single nucleotide polymorphism) panel analysis performed by The Jackson Laboratory revealed that this strain is on a mixed STOCK background (5 out of 22 markers are segregating for C57BL/6, FVB/N, or 129).
Expressed Gene | CYP1A2, cytochrome P450 family 1 subfamily A member 2, human |
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Expressed Gene | CYP1A1, cytochrome P450 family 1 subfamily A member 1, human |
Site of Expression |
Allele Name | targeted mutation 1, Daniel W Nebert |
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Allele Type | Targeted (Null/Knockout) |
Allele Synonym(s) | Cyp1a1/1a2-; Cyp1a2/Cyp1a1tm2Dwn; Del(9Cyp1a2-Cyp1a1)1Dwn |
Gene Symbol and Name | Del(9Cyp1a2-Cyp1a1)1Dwn, deletion, Chr 9, Daniel W Nebert 1 |
Gene Synonym(s) | |
Strain of Origin | 129P2/OlaHsd or 129S6/SvEvTac |
Chromosome | 9 |
Molecular Note | The Cyp1a2(t) targeted allele (J:122409) was generated using a targeting vector designed to insert a loxP-flanked PGK-NEO cassette 350 bp downstream of the endogenous stop codon (about 60 bp 3' of exon 7). The construct was electroporated into 129S6/SvEvTac-derived embryonic stem (ES) cells which were microinjection into C57BL/6 blastocysts. The chimeras were bred with C57BL/6 to generate Cyp1a2(t) mutant mice. To create the Cyp1a1 targeted allele (J:59398), a targeting vector was designed to insert a loxP-flanked hypoxanthine phosphoribosyltransferase (HPRT) minigene in intron 1 and a loxP site downstream of the termination codon in exon 7. Following electroporation into E14TG2a (HPRT-) ES cells and ES cell microinjection into the blastocoele cavity of C57BL/6J embryos, chimeric males were bred with C57BL/6J females. As described in J:86748, these Cyp1a1(t) mutant mice were then bred to a Cre-deleter strain (CAGGS-CRE, mixed C57BL/6J and FVB/NJ genetic background). The resulting transgenic mice found to be heterozygous for the floxed null Cyp1a1(-) allele (containing only exon 1, a portion of intron 1, and one remaining loxP site in the 3' UTR) were bred to mice heterozygous for the Cyp1a2(t) allele (as described in J:122409). Mutant mice (Cyp1a2(t), Cyp1a1(-), CAGGS-CRE) were bred to C57BL/6. Because of the close genomic position of these two genes, offspring having undergone Cre recombinase-mediated interchromosomal recombination between the loxP sites 3' beyond the stop codons of the Cyp1a2 and Cyp1a1 genes could be selected. Such mice were backcrossed to C57BL/6 for 10 generations (while selecting against the Cre-deleter transgene) to generate Cyp1a1/1a2 mutant mice. |
Allele Name | transgene insertion 1, Daniel W Nebert |
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Allele Type | Transgenic (Inserted expressed sequence, Humanized sequence) |
Allele Synonym(s) | hCYP1A1_1A2, BAC-H |
Gene Symbol and Name | Tg(CYP1A1,CYP1A2)1Dwn, transgene insertion 1, Daniel W Nebert |
Gene Synonym(s) | |
Promoter | CYP1A2, cytochrome P450 family 1 subfamily A member 2, human |
Promoter | CYP1A1, cytochrome P450 family 1 subfamily A member 1, human |
Expressed Gene | CYP1A2, cytochrome P450 family 1 subfamily A member 2, human |
Expressed Gene | CYP1A1, cytochrome P450 family 1 subfamily A member 1, human |
Strain of Origin | (C57BL/6J x DBA/2J)F1 |
Chromosome | UN |
Molecular Note | To create "humanized" CYP1A1 and CYP1A2 transgenic mice (hCYP1A1_1A2), a single copy of the 180-kb human CYP1A1_CYP1A2 locus-containing BAC-H (BAC Human CTB clone 31H21: including the 23.3 kb spacer region, 90 kb of CYP1A2 3'-flanking region and 53 kb of CYP1A1 3' flanking region) was microinjected into (C57BL/6J x DBA/2J)F1 oocytes. Founder mice having a single copy of BAC-H were identified and then backcrossed for 10 generations to C57BL/6 mice. |
When maintaining a live colony, these mice (homozygous for the Cyp1a1/1a2 targeted allele and hemizygous for the hCYP1A1_1A2 transgene) may be bred together.
When using the STOCK Cyp1atm1Dwn Tg(CYP1A1,CYP1A2)1Dwn/J mouse strain in a publication, please cite the originating article(s) and include JAX stock #007580 in your Materials and Methods section.
Facility Barrier Level Descriptions
Service/Product | Description | Price |
---|---|---|
Heterozygous for Cyp1a2, Hemizygous or Non carrier for Tg(CYP1A1,CYP1A2)1Dwn |
Frozen Mouse Embryo | STOCK Del(9Cyp1a2-Cyp1a1)1Dwn Tg(CYP1A1 CYP1A2)1Dwn/J Frozen | $2595.00 |
Frozen Mouse Embryo | STOCK Del(9Cyp1a2-Cyp1a1)1Dwn Tg(CYP1A1 CYP1A2)1Dwn/J Frozen | $2595.00 |
Frozen Mouse Embryo | STOCK Del(9Cyp1a2-Cyp1a1)1Dwn Tg(CYP1A1 CYP1A2)1Dwn/J Frozen | $3373.50 |
Frozen Mouse Embryo | STOCK Del(9Cyp1a2-Cyp1a1)1Dwn Tg(CYP1A1 CYP1A2)1Dwn/J Frozen | $3373.50 |
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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.
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