Construction and Biological Evaluation of a Novel Integrin ανβ₃-Specific Carrier for Targeted siRNA Delivery In Vitro.
(1) Background: The great potential of RNA interference (RNAi)-based gene therapy is premised on the effective delivery of small interfering RNAs (siRNAs) to target tissues and cells. Hence, we aimed at developing and examining a novel integrin αvβ₃-specific delivery carrier for targeted transfection of siRNA to malignant tumor cells; (2) Methods: Arginine-glycine-aspartate motif (RGD) was adopted as a tissue target for specific recognition of integrin αvβ₃. To enable siRNA binding, a chimeric peptide was synthesized by adding nonamer arginine residues (9R) at the carboxy terminus of cyclic-RGD dimer, designated as c(RGD)₂-9R. The efficiency of 9R peptide transferring siRNA was biologically evaluated in vitro by flow cytometry, confocal microscopy, and Western blot; (3) Results: An optimal 10:1 molar ratio of c(RGD)₂-9R to siRNA was confirmed by the electrophoresis on agarose gels. Both the flow cytometry and confocal microscopy results testified that transfection of c(RGD)₂-9R as an siRNA delivery carrier was obviously higher than the naked-siRNA group. The results of Western blot demonstrated that these 9R peptides were able to transduce siRNA to HepG2 cells in vitro, resulting in efficient gene silencing; and (4) Conclusion: The chimeric peptide of c(RGD)₂-9R can be developed as an effective siRNA delivery carrier and shows potential as a new strategy for RNAi-based gene therapy.
Department of Nuclear Medicine, Peking University First Hospital, No. 8, Xishiku St., West District, Beijing 100034, China. xueqichen@bjmu.edu.cn.
Xueqi
X
Chen
Department of Nuclear Medicine, Peking University First Hospital, No. 8, Xishiku St., West District, Beijing 100034, China. louisa_liu@bjmu.edu.cn.
Meng
M
Liu
Department of Nuclear Medicine, Peking University First Hospital, No. 8, Xishiku St., West District, Beijing 100034, China. rongfu_wang@163.com.
Rongfu
R
Wang
Department of Nuclear Medicine, Peking University First Hospital, No. 8, Xishiku St., West District, Beijing 100034, China. pingyanzheng@126.com.
Ping
P
Yan
Department of Nuclear Medicine, Peking University First Hospital, No. 8, Xishiku St., West District, Beijing 100034, China. zhangcl0326@163.com.
Chunli
C
Zhang
Department of Nuclear Medicine, Peking University First Hospital, No. 8, Xishiku St., West District, Beijing 100034, China. machao00225@163.com.
Chao
C
Ma
Department of Nuclear Medicine, Peking University First Hospital, No. 8, Xishiku St., West District, Beijing 100034, China. jack-yin@163.com.
Lei
L
Yin
D045744:Cell Line, Tumor; D002478:Cells, Cultured; D016503:Drug Delivery Systems; D005434:Flow Cytometry; D015870:Gene Expression; D020868:Gene Silencing; D018014:Gene Transfer Techniques; D006801:Humans; D066298:In Vitro Techniques; D039302:Integrin alphaVbeta3 / Q000737:chemistry*; D015394:Molecular Structure; D010455:Peptides / Q000737:chemistry; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000008:administration & dosage / Q000737:chemistry*; D014162:Transfection
D039302:Integrin alphaVbeta3; D010455:Peptides; D034741:RNA, Small Interfering
RNA interference (RNAi); arginine-glycine-aspartate (RGD); integrin αvβ3; small interfering RNA (siRNA)