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Gene silencing by RNA interference has become a very promising technique in the fields of biomedical and life sciences during the past few years. In this approach, small interfering RNA molecules that specifically bind to and cleave transcripts of target genes are introduced into cells, thereby causing full or partial loss of gene function, i.e. comprising null or hypomorphic phenotypes. That is why accession to and characterizations of genes that are associated with diseases and/or disease progression has become easier and faster in recent times. In this review the discovery of gene silencing by RNA interference, its molecular mechanism, optimal design principles of small interfering RNAs, advantages and barriers of RNA interference as a gene silencing tool, and finally its therapeutic applications with outcomes in some clinical trials with future aspects including oncolytic viruses will be discussed.