P2ry12^{em1(icre/ERT2)Tda}

The allele was created using CRISPR/cas9 endonuclease-mediated genome editing to target the stop codon of the P2ry12 locus on chromosome 3. The cas9-RNP complex and a 2A-iCreERT2 dsDNA (encoding a viral 2A oligopeptide that mediates ribosomal skipping and the iCreER fusion gene with ~1.5 kbp homology arms) were injected into the pronucleus of C57BL/6N mouse embryos, followed by adoptive embryo transfer. Founder mice and progeny were screened by PCR, and proper genomic integration was verified by Southern blot and genomic sequencing. Founders were bred to C57BL/6J mice for germline transmission. (J:291145)