B6;129-Hprt^{tm2(CMV-tdTomato)Nat}/J

Stock number: 021428
Research areas: Research Tools

Description

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.

Detailed description

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.

Development

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 Hprt^b-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.

Allele

Symbol: Hprt^{tm2(CMV-tdTomato)Nat}
Name: targeted mutation 2, Jeremy Nathans
MGI accession ID: MGI:5449929
Generation method: Targeted
Attributes: Conditional ready (e.g. floxed); No functional change
Marker symbol: Hprt
Marker name: hypoxanthine guanine phosphoribosyl transferase
Chromosome: X
Strain of origin: (B6.129P2-Hprt^b-m3/J x 129S-Gt(ROSA)26Sor^tm1Sor/J)F1
Expressed genes: RFP,Red Fluorescent Protein,coral
Site of expression: tdTomato fluorescent protein will be expressed in cells where promoters driving Cre recombinase are expressed.
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.