Functional comparison of single- and double-stranded siRNAs in mammalian cells.
The concept of small interfering RNA (siRNA) has been extended to include not only short double-stranded RNA of 19-25bp, but also single-stranded antisense RNA of the same length, since such single-stranded antisense siRNAs were recently found to be able to inhibit gene expression as well. We made comprehensive comparison of double- and single-stranded siRNA functions in RNA interference (RNAi), targeting multiple sites and different mRNAs, measuring RNAi effects at different time-points and in different cell lines, and examining response curves. Duplex siRNAs were found to be more potent than single-stranded antisense siRNAs. This was verified by the observation that single-stranded antisense siRNAs, which were inefficient in some cases when used alone, could be rescued from inefficiency by sequentially transfecting with the sense siRNAs. This result suggests that the structural character of siRNA molecules might be a more important determinant of siRNA efficiency than the cellular persistence of them.
Xu
Yunhe
Y
Center for Genomics and Bioinformatics, Karolinska Institute, SE-17177 Stockholm, Sweden. Yunhe.Xu@cgb.ki.se
Linde
Annika
A
Larsson
Ola
O
Thormeyer
Dorit
D
Elmen
Joacim
J
Wahlestedt
Claes
C
Liang
Zicai
Z
D000818:Animals; D015703:Antigens, CD / Q000096:biosynthesis / Q000235:genetics; D001665:Binding Sites; D002460:Cell Line; D002469:Cell Separation; D002478:Cells, Cultured; D005434:Flow Cytometry; D017930:Genes, Reporter; D006801:Humans; D008156:Luciferases / Q000378:metabolism; D051916:Membrane Cofactor Protein; D008562:Membrane Glycoproteins / Q000096:biosynthesis / Q000235:genetics; D051379:Mice; D008969:Molecular Sequence Data; D041681:NIH 3T3 Cells; D016376:Oligonucleotides, Antisense / Q000737:chemistry; D012313:RNA / Q000737:chemistry; D034622:RNA Interference; D012330:RNA, Double-Stranded / Q000737:chemistry*; D012333:RNA, Messenger / Q000378:metabolism; D034741:RNA, Small Interfering / Q000737:chemistry*; D014162:Transfection
D015703:Antigens, CD; C497857:CD46 protein, human; C497859:Mcp protein, mouse; D051916:Membrane Cofactor Protein; D008562:Membrane Glycoproteins; D016376:Oligonucleotides, Antisense; D012330:RNA, Double-Stranded; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D012313:RNA; D008156:Luciferases