The construction and identification of Fas-targeting siRNA-expressing plasmid / 西安交通大学学报(医学版)

ABSTRACT

Objective To construct Fas-targeting siRNA-expressing plasmid, and explore the significance of Fas inhibition in the treatment of acute aplastic anemia. Methods U6 promoter cassette and siFas sequence were obtained by PCR method, and cloned into modified pcDNA3.1 to produce plasmid pU6-siFas, which was transfected into P815 cells with limpofectin 2000, and then Fas expression was detected by immunohistochemistry. Results The plasmid pU6-siFas could efficiently reduce the expression of Fas and confer G-418 resistance in P815 cells. Conclusion The successful construction of the siRNA expressing plasmid will facilitate the application of RNA interference technique, and lay foundation for further studies of the therapeutic effect of Fas inhibition on acute aplastic anemia.