title: Silencing of BCR/ABL Chimeric Gene in Human Chronic Myelogenous Leukemia Cell Line K562 by siRNA-Nuclear Export Signal Peptide Conjugates.
abstract: Herein we described the synthesis of siRNA-NES (nuclear export signal) peptide conjugates by solid phase fragment coupling and the application of them to silencing of bcr/abl chimeric gene in human chronic myelogenous leukemia cell line K562. Two types of siRNA-NES conjugates were prepared, and both sense strands at 5' ends were covalently linked to a NES peptide derived from TFIIIA and HIV-1 REV, respectively. Significant enhancement of silencing efficiency was observed for both of them. siRNA-TFIIIA NES conjugate suppressed the expression of BCR/ABL gene to 8.3% at 200 nM and 11.6% at 50 nM, and siRNA-HIV-1REV NES conjugate suppressed to 4.0% at 200 nM and 6.3% at 50 nM, whereas native siRNA suppressed to 36.3% at 200 nM and 30.2% at 50 nM. We could also show complex of siRNA-NES conjugate and designed amphiphilic peptide peptideβ7 could be taken up into cells with no cytotoxicity and showed excellent silencing efficiency. We believe that the complex siRNA-NES conjugate and peptideβ7 is a promising candidate for in vivo use and therapeutic applications.
authors.affiliation: 1 Department of Biological and Environmental Chemistry, School of Humanity Oriented Science and Technology, Kindai University , Iizuka, Japan .
authors.forename: Yasuhiro
authors.initials: Y
authors.lastname: Shinkai
authors.affiliation: 1 Department of Biological and Environmental Chemistry, School of Humanity Oriented Science and Technology, Kindai University , Iizuka, Japan .
authors.forename: Shinichi
authors.initials: S
authors.lastname: Kashihara
authors.affiliation: 1 Department of Biological and Environmental Chemistry, School of Humanity Oriented Science and Technology, Kindai University , Iizuka, Japan .
authors.forename: Go
authors.initials: G
authors.lastname: Minematsu
authors.affiliation: 1 Department of Biological and Environmental Chemistry, School of Humanity Oriented Science and Technology, Kindai University , Iizuka, Japan .
authors.forename: Hirofumi
authors.initials: H
authors.lastname: Fujii
authors.affiliation: 1 Department of Biological and Environmental Chemistry, School of Humanity Oriented Science and Technology, Kindai University , Iizuka, Japan .
authors.forename: Madoka
authors.initials: M
authors.lastname: Naemura
authors.affiliation: 1 Department of Biological and Environmental Chemistry, School of Humanity Oriented Science and Technology, Kindai University , Iizuka, Japan .
authors.forename: Yojiro
authors.initials: Y
authors.lastname: Kotake
authors.affiliation: 1 Department of Biological and Environmental Chemistry, School of Humanity Oriented Science and Technology, Kindai University , Iizuka, Japan .
authors.forename: Yasutaka
authors.initials: Y
authors.lastname: Morita
authors.affiliation: 1 Department of Biological and Environmental Chemistry, School of Humanity Oriented Science and Technology, Kindai University , Iizuka, Japan .
authors.forename: Koichiro
authors.initials: K
authors.lastname: Ohnuki
authors.affiliation: 2 Institute of Chemical Biology and Fundamental Medicine , Siberian Branch of the Russian Academy of Sciences, Novosibirsk, Russia .
authors.forename: Alesya A
authors.initials: AA
authors.lastname: Fokina
authors.affiliation: 2 Institute of Chemical Biology and Fundamental Medicine , Siberian Branch of the Russian Academy of Sciences, Novosibirsk, Russia .|3 Novosibirsk State University , Novosibirsk, Russia .
authors.forename: Dmitry A
authors.initials: DA
authors.lastname: Stetsenko
authors.affiliation: 4 Institute of Fundamental Sciences, Massey University , Palmerston North, New Zealand .
authors.forename: Vyacheslav V
authors.initials: VV
authors.lastname: Filichev
authors.affiliation: 1 Department of Biological and Environmental Chemistry, School of Humanity Oriented Science and Technology, Kindai University , Iizuka, Japan .
authors.forename: Masayuki
authors.initials: M
authors.lastname: Fujii
mesh_terms: D065966:Absorption, Physicochemical; D004305:Dose-Response Relationship, Drug; D055808:Drug Discovery / Q000639:trends; D016044:Fusion Proteins, bcr-abl / Q000235:genetics; D015973:Gene Expression Regulation, Leukemic; D020868:Gene Silencing; D016313:Genes, abl / Q000187:drug effects* / Q000235:genetics; D006801:Humans; D020014:K562 Cells; D015464:Leukemia, Myelogenous, Chronic, BCR-ABL Positive / Q000235:genetics / Q000628:therapy*; D050997:Nanomedicine / Q000639:trends; D049790:Nuclear Export Signals / Q000235:genetics; D028861:Nuclear Pore Complex Proteins / Q000737:chemistry; D010446:Peptide Fragments / Q000138:chemical synthesis / Q000737:chemistry / Q000494:pharmacology*; D034741:RNA, Small Interfering / Q000737:chemistry / Q000235:genetics / Q000494:pharmacology*; D016601:RNA-Binding Proteins / Q000737:chemistry; D035582:Transcription Factor TFIIIA / Q000737:chemistry; D014162:Transfection
chemical_list: C119724:AGFG1 protein, human; D049790:Nuclear Export Signals; D028861:Nuclear Pore Complex Proteins; D010446:Peptide Fragments; D034741:RNA, Small Interfering; D016601:RNA-Binding Proteins; D035582:Transcription Factor TFIIIA; D016044:Fusion Proteins, bcr-abl
keywords: BCR/ABL; SPFC; siRNA-NES conjugates