Lack of interferon (IFN) response to T7 Transcribed pppG (n)(n = 2,3)-shRNA.
RNA interference (RNAi) mediated by siRNAs has proved to be a highly effective gene silencing mechanism with great potential for gene therapeutic applications. However, siRNA agents have been shown to exert non-target-related biological effects and toxicities, including immune stimulation. Specifically, siRNA synthesized from a T7 RNA polymerase system can trigger the potent induction of type I IFN in a variety of cells. The single-stranded RNA can also stimulate innate cytokine responses in mammals. We found that pppGn (n = 1-3), associated with the 5' end of the shRNA produced from the T7 RNA polymerase system, did not induce detectable levels of IFN. The residual amount of G associated with the 5'-end of the transcript was proportional to the reduction of the interferon response. We describe a T7 pppGn (n = 1-3) shRNA synthesis system that alleviates the IFN response, which will facilitate the design of siRNAs while maintaining their full efficacy.
Gondai
Takuma
T
Department of Life and Environmental Science, Chiba Institute of Technology, Narashino-shi, Chiba, Japan.
Yamaguchi
Kazuya
K
Hashimoto
Kahoko
K
Miyano-Kurosaki
Naoko
N
Takaku
Hiroshi
H
D001483:Base Sequence; D012321:DNA-Directed RNA Polymerases / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D007372:Interferons / Q000096:biosynthesis*; D008156:Luciferases; D008969:Molecular Sequence Data; D009841:Oligonucleotides / Q000235:genetics*; D012333:RNA, Messenger; D034741:RNA, Small Interfering / Q000235:genetics*; D014158:Transcription, Genetic; D014764:Viral Proteins / Q000378:metabolism*
D009841:Oligonucleotides; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D014764:Viral Proteins; D007372:Interferons; D008156:Luciferases; C062210:bacteriophage T7 RNA polymerase; D012321:DNA-Directed RNA Polymerases