Inhibition of C-raf expression by antisense oligonucleotides extends heart allograft survival in rats

Stanislaw M. Stepkowski, Xeimei Qu, Mou Er Wang, Ling Tian, Wenhao Chen, Edward V. Wancewicz, Joseph F. Johnston, C. Frank Bennett, Brett P. Monia

Research output: Contribution to journalArticle

21 Scopus citations

Abstract

Background. C-raf is a well-characterized serine/threonine (Ser/Thr) protein kinase that is involved in the transduction of multiple signals of T cells. We demonstrate that the inhibition of C-raf mRNA expression prolongs heart allograft survival. Methods. Three 20-mer C-raf antisense oligonucleotides, each with identical sequences, were synthesized with different chemical modifications: one as a uniform phosphorothioate oligodeoxynucleotide (PS oligo), a second with a PS backbone and 2'-methoxy-ethyl (ME) substitutions at the 2'-sugar positions in the first and last five nucleotides, and a third with a mixed PS and phosphodiester (PD) backbone and ME modifications on the first and last five nucleotides. Results. Both ME-modified C-raf antisense oligos were at least 5-fold more effective than the PS C-raf antisense oligo in blocking C-raf mRNA expression in two cell lines. Similarly, each of the ME C-raf antisense oligos produced better heart allograft survival rates than did PS C-raf oligo. Furthermore, although the combination of PS C-raf antisense oligo with sirolimus (SRL) acted synergistically to extend heart allograft survival, the effect was potentiated by either of the ME-modified oligos. Conclusions. C-raf inhibition extends heart allograft survival, and ME-modification potentiates antisense activity.

Original languageEnglish (US)
Pages (from-to)656-661
Number of pages6
JournalTransplantation
Volume70
Issue number4
DOIs
StatePublished - Aug 27 2000

ASJC Scopus subject areas

  • Transplantation

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