B6;129-Gt(ROSA)26Sor^tm2Nat/J

Stock number: 009253
Product name: ROSA26-AP Inversion (R26iAP)
Research areas: Research Tools

Description

This strain carries an inverted human placental alkaline phosphatase (ALPP; PLAP) coding segment flanked by oppositely-oriented loxP sites integrated in the Gt(ROSA)26Sor gene. Cre excision of the floxed segment flips rather than excises the ALPP inversion and reinstates a transcriptionally active form of the gene. This reporter can be activated by tissue-specific cre recombinase in any cell in the body at any time in development. The result is very clear ALPP enzyme activity that can be detected histochemically.

Detailed description

This strain carries an inverted human placental alkaline phosphatase (ALPP; PLAP) coding segment flanked by oppositely-oriented loxP sites integrated in the Gt(ROSA)26Sor gene. Cre excision of the floxed segment flips rather than excises the ALPP inversion and reinstates a transcriptionally active form of the gene. This reporter can be activated by tissue-specific cre recombinase in any cell in the body at any time in development. The result is very clear ALPP enzyme activity that can be detected histochemically. Heterozygotes and homozygotes are normal in size, viability, and fertility.

Development

The human placental alkaline phosphatase (ALPP; PLAP) coding region was engineered to contain a single intron with a loxP site. Another loxP site in inverted orientation was placed 3' of the ALPP coding region. A single round of Cre-mediated recombination in E. coli resulted in the inversion of the 3' half of the coding region, thereby disrupting the ALPP open reading frame. A PGK-neo site is located downstream of the 3' UTR. This construct, referred to as "AP inversion" or "AP inv", was knocked into the ubiquitously-expressed Gt(ROSA)26Sor locus by homologous recombination. A 129-derived embryonic stem cell line was used to create the mutation. This line was backcrossed three times to C57BL/6 by the donating laboratory.

Allele

Symbol: Gt(ROSA)26Sor^tm2Nat
Name: targeted mutation 2, Jeremy Nathans
MGI accession ID: MGI:3834904
Generation method: Targeted
Attributes: Reporter
Marker symbol: Gt(ROSA)26Sor
Marker name: gene trap ROSA 26, Philippe Soriano
Chromosome: 6
Strain of origin: 129
Expressed genes: ALPP,alkaline phosphatase, placental,human
Molecular note: The human placental alkaline phosphatase (ALPP; PLAP) coding region was engineered to contain a single intron with a loxP site. Another loxP site in inverted orientation was placed 3' of the ALPP coding region. A single round of Cre-mediated recombination in E. coli resulted in the inversion of the 3' half of the coding region, thereby disrupting the ALPP open reading frame. This construct, referred to as "AP inversion" or "AP inv", was knocked into the ubiquitously-expressed Gt(ROSA)26Sor locus by homologous recombination.