Antisense Sites Screening of Fas Gene mRNA and Its Validation in vitro / 中国生物工程杂志

ABSTRACT

Three candidate antisense target sites of mouse Fas gene were screened by PARASS (poly-A anchored RNA accessible sites screening) technology. They were target at Fas gene 297nt-317nt, 618nt- 638nt and 662nt-682nt. Antisense oligos (A1, A2 and A3) and DNAzymes (D1, D2, and D3) for every target site were designed and synthesized. In vitro, the validation of the sites were judged by antisense oligos included RNase H splicing and the DNAzyme degradation. The results indicated that A1, A2 and A3 introduced RNase H degradation. DNAzymes D1, D2 and D3 cleaved Fas mRNA effectively. Neither degradation observed in antisense oligo RNase H group in non-target site (1211-1231nt) and 2 bases mismatched of A3, nor splicing occurred in DNzyme group in non-target site ( 1211-1231nt) and 2 bases mismatched of D3. Site 2 and 3 were at the same positions with those of ISIS Pharmaceuticals. The effective antisense oligos and DNAzymes for Fas gene could be used for the research subsequently.