title: Amino-modified and lipid-conjugated dicer-substrate siRNA enhances RNAi efficacy.
abstract: The development of Dicer-substrate small interfering RNAs (DsiRNAs) has been pursued in recent years because these molecules exhibit a much more potent gene-silencing effect than 21-nucleotide (nt) siRNAs. In the present study, we designed eight different types of amino-modified DsiRNAs and a palmitic acid-conjugated DsiRNA expected to result in improved biological properties of siRNAs, including their stability against nuclease degradation, membrane permeability, and RNAi efficacy. The DsiRNAs were modified with an amine at the 5'- and/or 3'-end of the sense and/or antisense strand. Dicer enzyme cleaved most of the amino-modified DsiRNAs to lead to the release of 21-nt siRNA; some of them, however, were not or partly cleaved. All amino-modified DsiRNAs exhibited strong resistance against nuclease degradations. Among the amino-modified DsiRNAs, the DsiRNA modified with an amine restricted at the 3'-end of the sense strand showed the most enhanced gene-silencing effect and maintained its potent gene suppression after one week of cell transfection against Renilla luciferase activity. For further improvement, palmitic acid was conjugated to DsiRNA at the 3'-end of the sense strand (C16-DsiRNA) to facilitate the membrane permeability and potent gene-silencing activity. The C16-DsiRNA showed enhanced membrane permeability to HeLa cells. The C16-DsiRNA exhibited extremely high inhibition of Renilla luciferase activity.
authors.affiliation: Laboratory of Molecular Cell Biology, Department of Life Science, Yasuda Women's University Faculty of Pharmacy, Hiroshima, Japan. kubot@yasuda-u.ac.jp
authors.forename: Takanori
authors.initials: T
authors.lastname: Kubo
authors.forename: Yoshifumi
authors.initials: Y
authors.lastname: Takei
authors.forename: Keichiro
authors.initials: K
authors.lastname: Mihara
authors.forename: Kazuyoshi
authors.initials: K
authors.lastname: Yanagihara
authors.forename: Toshio
authors.initials: T
authors.lastname: Seyama
mesh_terms: D000588:Amines / Q000737:chemistry*; D002478:Cells, Cultured; D020868:Gene Silencing; D006367:HeLa Cells; D006801:Humans; D008055:Lipids / Q000737:chemistry*; D049410:Luciferases, Renilla / Q000096:biosynthesis / Q000235:genetics; D019308:Palmitic Acid / Q000737:chemistry; D034741:RNA, Small Interfering / Q000097:blood / Q000235:genetics* / Q000378:metabolism*; D043244:Ribonuclease III / Q000378:metabolism*; D013329:Structure-Activity Relationship
chemical_list: D000588:Amines; D008055:Lipids; D034741:RNA, Small Interfering; D019308:Palmitic Acid; D049410:Luciferases, Renilla; D043244:Ribonuclease III