title: Molecular Properties, Functional Mechanisms, and Applications of Sliced siRNA.
abstract: Using pre-miR-451 as a model molecule, we have characterized the general molecular properties of small hairpin RNAs that are processed into potent small interfering RNAs (siRNA) by Argonaute2 (Ago2). The Ago2-sliced siRNAs (sli-siRNAs) have the same silencing potency as the classical Dicer diced siRNAs (di-siRNAs) but have dramatically reduced unwanted sense strand activities. We have built vectors with the constitutive or inducible U6 promoter that can express sli-siRNAs in mammalian cells, in which the sli-siRNAs can be correctly processed to repress target genes. As a proof of principle for potential applications of sli-siRNAs in vivo, we show that the expression of one Ago2 shRNA-1148 in HCT-116 colon cancer cells knocked down RRM2 expression and reduced the proliferation and invasiveness of the cells. The defined sli-siRNA model molecules and the expression systems established in this study will facilitate the design and application of sli-siRNAs as novel potent RNAi triggers with reduced off-target effects.
authors.affiliation: Department of Molecular Pharmacology, Beckman Research Institute of the City of Hope, Duarte, California, USA.
authors.forename: Guihua
authors.initials: G
authors.lastname: Sun
authors.affiliation: Summer Interns, Beckman Research Institute of the City of Hope, Duarte, California, USA.
authors.forename: Spencer Yele
authors.initials: SY
authors.lastname: Yeh
authors.affiliation: Summer Interns, Beckman Research Institute of the City of Hope, Duarte, California, USA.
authors.forename: Christine Wan-Yin
authors.initials: CW
authors.lastname: Yuan
authors.affiliation: Summer Interns, Beckman Research Institute of the City of Hope, Duarte, California, USA.
authors.forename: Margaret Jean-Yih
authors.initials: MJ
authors.lastname: Chiu
authors.affiliation: Summer Interns, Beckman Research Institute of the City of Hope, Duarte, California, USA.
authors.forename: Bryan Shing Hei
authors.initials: BS
authors.lastname: Yung
authors.affiliation: Department of Molecular Pharmacology, Beckman Research Institute of the City of Hope, Duarte, California, USA.
authors.forename: Yun
authors.initials: Y
authors.lastname: Yen