title: A cytoplasmic pathway for gapmer antisense oligonucleotide-mediated gene silencing in mammalian cells.
abstract: Antisense oligonucleotides (ASOs) are known to trigger mRNA degradation in the nucleus via an RNase H-dependent mechanism. We have now identified a putative cytoplasmic mechanism through which ASO gapmers silence their targets when transfected or delivered gymnotically (i.e. in the absence of any transfection reagent). We have shown that the ASO gapmers can interact with the Ago-2 PAZ domain and can localize into GW-182 mRNA-degradation bodies (GW-bodies). The degradation products of the targeted mRNA, however, are not generated by Ago-2-directed cleavage. The apparent identification of a cytoplasmic pathway complements the previously known nuclear activity of ASOs and concurrently suggests that nuclear localization is not an absolute requirement for gene silencing.
authors.affiliation: Department of Medical Oncology, City of Hope, 1500 East Duarte Road, Duarte CA 91010, USA cstein@coh.org.
authors.forename: Daniela
authors.initials: D
authors.lastname: Castanotto
authors.affiliation: Department of Cancer Immunotherapeutics and Tumor Immunology, City of Hope, 1500 East Duarte Road, Duarte CA 91010, USA.
authors.forename: Min
authors.initials: M
authors.lastname: Lin
authors.affiliation: Department of Molecular Medicine, City of Hope, 1500 East Duarte Road, Duarte CA 91010, USA.
authors.forename: Claudia
authors.initials: C
authors.lastname: Kowolik
authors.affiliation: Pfizer Research Technology Center, 620 Memorial Drive, Cambridge, MA 02139, USA.
authors.forename: LiAnn
authors.initials: L
authors.lastname: Wang
authors.affiliation: Pfizer Research Technology Center, 620 Memorial Drive, Cambridge, MA 02139, USA.
authors.forename: Xiao-Qin
authors.initials: XQ
authors.lastname: Ren
authors.affiliation: bioTheranostics, 9640 Towne Center Dr., Suite 100, San Diego, CA 92121, USA.
authors.forename: Harris S
authors.initials: HS
authors.lastname: Soifer
authors.affiliation: Roche, Fremtidsvej 3, Horsholm, DK 2970, Denmark.
authors.forename: Troels
authors.initials: T
authors.lastname: Koch
authors.affiliation: Roche, Fremtidsvej 3, Horsholm, DK 2970, Denmark.
authors.forename: Bo Rode
authors.initials: BR
authors.lastname: Hansen
authors.affiliation: Roche, Fremtidsvej 3, Horsholm, DK 2970, Denmark.
authors.forename: Henrik
authors.initials: H
authors.lastname: Oerum
authors.affiliation: Department of Neuroscience, City of Hope, 1500 East Duarte Road, Duarte CA 91010, USA.
authors.forename: Brian
authors.initials: B
authors.lastname: Armstrong
authors.affiliation: Pfizer Research Technology Center, 620 Memorial Drive, Cambridge, MA 02139, USA.
authors.forename: Zhigang
authors.initials: Z
authors.lastname: Wang
authors.affiliation: Pfizer Research Technology Center, 620 Memorial Drive, Cambridge, MA 02139, USA.
authors.forename: Paul
authors.initials: P
authors.lastname: Bauer
authors.affiliation: Department of Molecular and Cellular Biology, City of Hope, 1500 East Duarte Road, Duarte CA 91010, USA.
authors.forename: John
authors.initials: J
authors.lastname: Rossi
authors.affiliation: Department of Medical Oncology, City of Hope, 1500 East Duarte Road, Duarte CA 91010, USA dcastanotto@coh.org.
authors.forename: C A
authors.initials: CA
authors.lastname: Stein
mesh_terms: D060565:Argonaute Proteins / Q000378:metabolism; D002460:Cell Line; D003593:Cytoplasm / Q000737:chemistry / Q000378:metabolism*; D020868:Gene Silencing; D018014:Gene Transfer Techniques; D016376:Oligonucleotides, Antisense / Q000032:analysis / Q000737:chemistry / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D034741:RNA, Small Interfering; D014162:Transfection
chemical_list: D060565:Argonaute Proteins; D016376:Oligonucleotides, Antisense; D012333:RNA, Messenger; D034741:RNA, Small Interfering