tailieunhanh - Báo cáo khoa học: A new and efficient method for inhibition of RNA viruses by DNA interference

We report here a new method for inhibition of RNA viruses induced by dsDNA. We demonstrated that both long dsDNA molecules and short interfering DNA with a sequence complementary to that of viral RNA inhibited tobacco mosaic virus expression and prevented virus spread. Also, the expression of the HIV-1 gp41 gene in HeLa cells was inhibited by com-plementary short interfering DNA. | ễFEBS Journal A new and efficient method for inhibition of RNA viruses by DNA interference Monika Nowak1 Eliza Wyszko1 Agnieszka Fedoruk-Wyszomirska1 Henryk Pospieszny2 Mirostawa Z. Barciszewska1 and Jan Barciszewski1 1 Institute of Bioorganic Chemistry Polish Academy of Sciences Poznan Poland 2 Department of Virology and Bacteriology Institute of Plant Protection Poznan Poland Keywords Dicer DNA interference gene silencing RNA interference siDNA Correspondence J. Barciszewski Institute of Bioorganic Chemistry of the Polish Academy of Sciences Noskowskiego 12 61-704 Poznan Poland Fax. 48 61 8520532 Tel 48 61 8528503 E-mail Received 16 February 2009 revised 2 June 2009 accepted 10 June 2009 doi We report here a new method for inhibition of RNA viruses induced by dsDNA. We demonstrated that both long dsDNA molecules and short interfering DNA with a sequence complementary to that of viral RNA inhibited tobacco mosaic virus expression and prevented virus spread. Also the expression of the HIV-1 gp41 gene in HeLa cells was inhibited by complementary short interfering DNA. We showed that Dicer processed dsDNA which suggests activation of the cellular machinery involved in silencing of RNA. For the silencing of viral RNA effected with dsDNA we coined the term DNA interference technology. Introduction RNA technologies which began 30 years ago with the antisense oligonucleotides and progressed through ribozymes and DNAzymes deoxyribozymes and their analogues have not met expectations as they have failed to deliver a suitable agent that can effectively inhibit gene expression at the RNA level. RNA interference RNAi technology is the most recent in the long line of nucleic acid-based therapeutic candidates. RNAi is induced by long dsRNA processed by the endonuclease Dicer into 21 nucleotide short interfering RNAs siRNAs or other 19-28 nucleotide small RNAs sRNAs 1 2 . These short stretches of RNA with 3 .