title: Antisense oligonucleotides capable of promoting specific target mRNA reduction via competing RNase H1-dependent and independent mechanisms.
abstract: Antisense oligonucleotides (ASOs) are most commonly designed to reduce targeted RNA via RNase H1-dependent degradation. In this paper we demonstrate that cellular proteins can compete for sites targeted by RNase H1-dependent ASOs. We further show that some ASOs designed to mediate RNase H1 cleavage can, in certain instances, promote target reduction both by RNase H1-mediated cleavage and by steric inhibition of binding of splicing factors at a site required for efficient processing of the pre-mRNA. In the latter case, RNase H cleavage was prevented by binding of a second protein, HSPA8, to the ASO/pre-mRNA heteroduplex. In addition, using a precisely controlled minigene system, we directly demonstrated that activity of ASOs targeting sites in introns is strongly influenced by splicing efficiency.
authors.affiliation: Department of Core Antisense Research, ISIS Pharmaceuticals, Inc., Carlsbad, California, United States of America.
authors.forename: Timothy A
authors.initials: TA
authors.lastname: Vickers
authors.affiliation: Department of Core Antisense Research, ISIS Pharmaceuticals, Inc., Carlsbad, California, United States of America.
authors.forename: Stanley T
authors.initials: ST
authors.lastname: Crooke
mesh_terms: D045744:Cell Line, Tumor; D006367:HeLa Cells; D006801:Humans; D007438:Introns / Q000235:genetics; D016376:Oligonucleotides, Antisense / Q000235:genetics*; D012322:RNA Precursors / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics*; D016914:Ribonuclease H / Q000378:metabolism*
chemical_list: D016376:Oligonucleotides, Antisense; D012322:RNA Precursors; D012333:RNA, Messenger; D016914:Ribonuclease H; C078856:ribonuclease HI