both strands of sirna have potential to guide posttranscriptional gene silencing in mammalian cells两股sirna潜在指导转录后的基因沉默在哺乳动物细胞.pdfVIP

Both Strands of siRNA Have Potential to Guide Posttranscriptional Gene Silencing in Mammalian Cells 1,2. 1,3. 3. 1 4 2 1 Jun-Xia Wei , Jie Yang , Ji-Feng Sun , Lin-Tao Jia , Yong Zhang , Hui-Zhong Zhang , Xia Li , Yan- 4 1 1,4 Ling Meng , Li-Bo Yao , An-Gang Yang * 1 State Key Laboratory of Cancer Biology; Department of Biochemistry and Molecular Biology, Fourth Military Medical University, Xi’an, China, 2 Department of Clinical Diagnosis, Tangdu Hospital, Fourth Military Medical University, Xi’an, China, 3 Department of Nephrology, Tangdu Hospital, Fourth Military Medical University, Xi’an, China, 4 Department of Immunology, Fourth Military Medical University, Xi’an, China Abstract Despite the widespread application of RNA interference (RNAi) as a research tool for diverse purposes, the key step of strand selection of siRNAs during the formation of RNA-induced silencing complex (RISC) remains poorly understood. Here, using siRNAs targeted to the complementary region of Survivin and the effector protease receptor 1 (EPR-1), we show that both strands of the siRNA duplex can find their target mRNA and are equally eligible for assembly into Argonaute 2 (Ago2) of RISC in HEK293 cells. Transfection of the synthetic siRNA duplexes with different thermodynamic profiles or short hairpin RNA (shRNA) vectors that generate double-stranded RNAs (dsRNAs), permitting processing specifically from either the 59 or 39 end of the incipient siRNA, results in the degradation of the respective target mRNAs of either strand of the siRNA duplex with comparable efficiencies. Thus, while most RNAi reactions may follow the thermodynamic asymmetry rule in strand