Cre excision of a floxed stop sequence enables CMV-directed expression of nuclear localized (nls) tdTomato (tdT) fluorescent protein in this Hprt (hypoxanthine guanine phosphoribosyl transferase) knock-in line.
Jeremy Nathans, Johns Hopkins University
Genetic Background | Generation |
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|
Allele Type | Gene Symbol | Gene Name |
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Targeted (Conditional ready (e.g. floxed), No functional change) | Hprt | hypoxanthine guanine phosphoribosyl transferase |
This X-linked knock-in of the Hprt (hypoxanthine guanine phosphoribosyl transferase) gene gives no expression of the nuclear localized (nls) tdTomato (tdT) fluorescent reporter in the absence of Cre-mediated recombination. Following excision of a floxed stop sequence, the nls-tdT coding region is expressed, driven by a CMV promoter. This reporter can be activated by Cre recombinase in any cell in the body at any time in development. In females, it is subject to X-inactivation.
A pair of insulator sequences, CMV enhancer/beta-actin promoter, loxP site, 3 stop transcription sites, loxP site, nuclear localized (nls) tdT-3xHA (tdTomato-hemagglutinin) epitope, rabbit beta-globin 3' UTR, and a pair of insulator sequences were inserted into the Hprt gene in the indicated order via targeted recombination. The backbone of the targeting vector was constructed by Dominic Ciavatta (Proc Natl Acad Sci U S A. 2006 Jun 27;103(26):9958-63. PMID 16777957). The mEMS1204 embryonic stem (ES) cell line, derived from B6;129 background mice harboring the Hprtb-m3 allele, was used to create the mutation (Genomics. 2009 Mar;93(3):196-204. PMID 18950699). The strain was maintained on a mixed B6 and 129 genetic background by the donating lab.
Expressed Gene | RFP, Red Fluorescent Protein, coral |
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Site of Expression | tdTomato fluorescent protein will be expressed in cells where promoters driving Cre recombinase are expressed. |
Allele Name | targeted mutation 2, Jeremy Nathans |
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Allele Type | Targeted (Conditional ready (e.g. floxed), No functional change) |
Allele Synonym(s) | Hprttm1(CMV-LSL-nls-tdT)Nat |
Gene Symbol and Name | Hprt, hypoxanthine guanine phosphoribosyl transferase |
Gene Synonym(s) | |
Promoter | CMV, cytomegalovirus, human |
Expressed Gene | RFP, Red Fluorescent Protein, coral |
Site of Expression | tdTomato fluorescent protein will be expressed in cells where promoters driving Cre recombinase are expressed. |
Strain of Origin | (B6.129P2-Hprtb-m3/J x 129S-Gt(ROSA)26Sortm1Sor/J)F1 |
Chromosome | X |
Molecular Note | A pair of insulator sequences, CMV enhancer/beta-actin promoter, loxP site, 3 stop transcription sites, loxP site, nuclear localized (nls) tdTomato-3xHA (tdTomato-hemagglutinin) epitope, rabbit beta-globin 3' UTR, and a pair of insulator sequences were inserted into the gene in the indicated order via targeted recombination. |
Homozygous females and hemizygous males are viable and fertile. This allele is subject to X-inactivation in females.
When using the B6;129-Hprttm2(CMV-tdTomato)Nat/J mouse strain in a publication, please cite the originating article(s) and include JAX stock #021428 in your Materials and Methods section.
Facility Barrier Level Descriptions
Service/Product | Description | Price |
---|---|---|
X linked -Heterozygous females and wildtype males for Hprt<tm2(CMV-tdTomato)Nat> |
Frozen Mouse Embryo | B6;129-Hprt<tm2(CMV-tdTomato)Nat>/J Frozen Embryo | $2595.00 |
Frozen Mouse Embryo | B6;129-Hprt<tm2(CMV-tdTomato)Nat>/J Frozen Embryo | $2595.00 |
Frozen Mouse Embryo | B6;129-Hprt<tm2(CMV-tdTomato)Nat>/J Frozen Embryo | $3373.50 |
Frozen Mouse Embryo | B6;129-Hprt<tm2(CMV-tdTomato)Nat>/J Frozen Embryo | $3373.50 |
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