The Role of RNAi in Mammalian Cells in Response to Sindbis virus infection

摘要: Abstract Viruses are obligate intracellular parasites that need to interact with their host in order replicate successfully. The understanding of this complex interaction between and virus is essential for developing new therapeutic strategies as viral infections pose a serious challenge healthcare, also because viruses impose enormous costs on the economy. This study focused role RNA interference an (Sindbis virus) its mammalian cell. A sensitive image–based replication assay was developed follow Sindbis HEK293C cells main anti-viral innate responses infection described. Virus increased Dicer helicase A (RHA) knockout cells, but not when central regulator interferon synthesis IRF3 knocked out. High-throughput Solexa Illumina sequencing used detect small (svRNA) (SINV) infected human changes cellular microRNA (miRNA) expression profile. Very few vsRNA sequences were detected by during infection, I argue they random degradation products, Dicer-generated svRNAs. Due very low level svRNAs, these undetectable using northern blotting. We have found profile miRNAs did change early stages according data, finding which verified functional RNAi assess activity function system subjected stress, type interferon, dsRNA, blotting verify data. certain stress signals -double stranded SINV infection- decrease efficiency siRNA knockdowns siRNA-based knockdown system. I identified two factors important (Dicer, RHA). lack fragments led conclusion pathway suppressed either cell or itself, although has been shown any suppressors previous studies conducted insect vector. can be explained fact both immunity same molecule, placing systems into direct competition substrate. My hypothesis so does process long dsRNA small, 21nt fragments, invisible immune system.