B6;129-Piezo1^tm1.1Apat/J

Stock number: 029214
Product name: Piezo1^P1-tdT
Research areas: Neurobiology Research, Research Tools

Description

These mice express a fluorescent tdTomato reporter from the Piezo1 promoter, making them suitable for applications related to vascular development and red blood cell volume.

Detailed description

Piezo1 (piezo-type mechanosensitive ion channel component 1), expressed in endothelial cells of developing blood vessels in mice, is required for vascular development. It encodes a mechanosensitive non-selective cation channel that, when activated, plays an essential role in red blood cell (RBC) volume homeostasis.

These mice express a fluorescent tdTomato reporter from the Piezo1 promoter. RT-PCR confirms the presence of a fusion product and qPCR analysis demonstrates protein levels similar to those of wildtype controls in endothelial cells of lung, kidney, spleen, heart, and liver. Both peripheral blood RBCs and developing bone marrow pro-RBCs exhibit increased tdTomato fluorescence by flow cytometry, as compared to wildtype mice. Western blot of peripheral RBCs shows clear expression of a ~320 kDA Piezo1-tdTomato fusion protein.

Development

TdTomato and a self-excising pACN neomycin cassette were inserted just before the endogenous Piezo1 stop codon in exon 51. The mutation was created via homologous recombination in 129/SvJ-derived embryonic stem cells. Cre recombinase elements within pACN cassette excised the neomycin feature. This strain has been backcrossed to C57BL/6J for more than 8 generation by the donating lab (see SNP note below). Agouti/brown coat color alleles may still be segregating.

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. Four of the 27 markers throughout the genome were segragating, suggesting an incomplete backcross. Also, 1 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 mixed C57BL/6J ; C57BL/6N genetic background.

Allele

Symbol: Piezo1^tm1.1Apat
Name: targeted mutation 1.1, Ardem Patapoutian
MGI accession ID: MGI:5762891
Generation method: Targeted
Attributes: Reporter
Marker symbol: Piezo1
Marker name: piezo-type mechanosensitive ion channel component 1
Chromosome: 8
Strain of origin: 129
Expressed genes: RFP,Red Fluorescent Protein,coral
Site of expression: tdTomato fluorescence is expressed in peripheral blood RBCs and developing bone marrow pro-red blood cells.
Molecular note: A tdTomato reporter gene and floxed, self-excising neomycin resistance cassette replaced the 3'UTR. The final allele lacks the neomycin resistance cassette.