Abstract: The aim of this study was to obtain the transient receptor potential M2 (TRPM2) gene knockout heterozygous mouse model by using CRISPR/cas9 technology. Referring to intron 2 and intron 24 sequences of Trpm2-203 in Ensembl database, sgRNA targets were selected and synthesized in vitro. Cas9 mRNA was obtained by in vitro transcription. Cas9 mRNA and sgRNA were microinjected into the fertilized eggs of C57BL/6J mice and transplanted into the fallopian tube of pseudopregnant female mice to obtain F0mice. Positive F0generation mice were mated with wild-type C57BL/6J mice to obtain F1generation. Three positive F0generation mice were obtained by sequencing PCR products. Four strains were obtained by mating between positive F0generation mice and wild-type C57BL/6J mice with a total of 27 positive F1generation mice. Verified by real time fluorescent quantitative PCR and Western Blot, it was shown that the TRPM2 knockout heterozygous mice (TRPM2+/-) were successfully constructed in this study, which provides a good animal model for the further study of the biological function of TRPM2 gene and its expression products.

Key words: CRISPR/Cas9, TRPM2, gene knockout, mice