TY - JOUR

T1 - Silencing of BCR/ABL Chimeric Gene in Human Chronic Myelogenous Leukemia Cell Line K562 by siRNA-Nuclear Export Signal Peptide Conjugates

AU - Shinkai, Yasuhiro

AU - Kashihara, Shinichi

AU - Minematsu, Go

AU - Fujii, Hirofumi

AU - Naemura, Madoka

AU - Kotake, Yojiro

AU - Morita, Yasutaka

AU - Ohnuki, Koichiro

AU - Fokina, Alesya A.

AU - Stetsenko, Dmitry A.

AU - Filichev, Vyacheslav V.

AU - Fujii, Masayuki

N1 - Funding Information: This work was financially supported by JSPS KAKEN grant number 22550159. D.A.S. is supported by RSF grant no. 14-44-00068. A.A.F. acknowledges support from RFBR grant no. 16-03-01055. Publisher Copyright: © 2017, Mary Ann Liebert, Inc. Copyright: Copyright 2018 Elsevier B.V., All rights reserved.

PY - 2017/6

Y1 - 2017/6

N2 - 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.

AB - 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.

KW - BCR/ABL

KW - SiRNA-NES conjugates

KW - SPFC

KW - siRNA-NES conjugates

KW - TRANSCRIPTION FACTOR

KW - OLIGONUCLEOTIDES

UR - http://www.scopus.com/inward/record.url?scp=85020229513&partnerID=8YFLogxK

U2 - 10.1089/nat.2016.0647

DO - 10.1089/nat.2016.0647

M3 - Article

C2 - 28355131

AN - SCOPUS:85020229513

VL - 27

SP - 168

EP - 175

JO - Nucleic Acid Therapeutics

JF - Nucleic Acid Therapeutics

SN - 2159-3337

IS - 3

ER -