Abstract: Aim To establish the stable inhibition of DAPK gene expression PC12 cell lines and observate the character Methods 5 pairs of shRNA were designed by using web soft ware, then they were synthesized and inserted into the pDsRed1-N1-U6 vector. The recombinant plasmids were transfected into PC12 cell and screening the monoclone by G418 and establishing the stable PC12 cell lines. DAPK expression was detected by Western blot. MTT, FCM assay were used to assess the cell lines growth. Results The recombinant plasmids were successfully constructed and transfected into PC12 cell, Western blot showed that the RNA interference effect of the pDsRed1-N1-U6-F2 was the best among them. Conclusion It is a feasible way to establish the stable inhibition of DAPK gene expression PC12 cell lines by shRNA expression vectors.

Key words: DAPK, RNAi, shRNA expression vectors, PC12 cell