P2ry1^em1Robn

Sequence coding for an HA tag was inserted immediately after the start codon and all six serine (TCC, AGT, TCC, AGT, TCT, AGT) and two threonine (ACT, ACG) codons within the sequence coding for the C-terminal tail were mutated to alanine codons (GCC, GCT, GCC, GCT, GCT, GCT) (GCT, GCG) (p.S343A, p.S346A, p.S352A, p.S354A, p.T358A, p.S363A, p.T371_S372delinsAA) using sgRNAs (targeting GGAAAGAAGAGGATGACCG and TGGAGACACGAGTTTGTGA) and ssODN templates with CRISPR/Cas9 technology. The mutations (SRRSEANLOSKSEDMTLNILSEFKONGDTSL to ARRAEANLQAKAEDMALNILAEFKQNGDAAL) block phosphorylation of the encoded residues and prevent desensitization of the encoded peptide in platelets. (J:342773)