期刊
JOURNAL OF GENE MEDICINE
卷 11, 期 9, 页码 804-812出版社
WILEY
DOI: 10.1002/jgm.1365
关键词
double-stranded RNA binding domain; human dsRNA activated protein kinase R; protein-mediated siRNA delivery; siRNA delivery
资金
- Ministry of Education, Science and Technology [M10414030002-05N1403-0210, R31-2008-000-10071-0]
Background A variety of synthetic carriers, such as cationic polymers and lipids, have been used as nonviral carriers for small interfering RNA (siRNA) delivery. Although siRNA polyplexes and lipoplexes exhibited good gene silencing efficiencies, they often showed serious cytotoxicities, which are not useful for clinical applications. A double-stranded RNA binding cellular protein with highly specific siRNA binding property and noncytotoxicity was used for siRNA delivery. Methods A double-stranded RNA binding domain (dsRBD) of human double-stranded RNA activated protein kinase R was genetically produced and utilized to complex siRNA for intracellular delivery. For characterization of the siRNA/dsRBD complexes, decomplexation assay and RNase protection assay were performed. Cytotoxicity and target gene inhibition ability were also examined using human carcinoma cell lines. Results The recombinantly produced polypeptide dsRBD exhibited its inherent binding activity for siRNA without sequence specificity, and the siRNA/dsRBD complexes protected siRNA from degradation by ribonucleases. Green fluorescent protein (GFP) siRNA/dsRBD complexes showed prominent down-regulation of a target GFP gene, when an endosomal escape function was supplemented by addition of a fusogenic peptide, KALA, in the formulation. Conclusions The results suggest that dsRBD-based protein carriers could be successfully applied for a wide range of therapeutic siRNAS for intracellular gene inhibition without showing any cytotoxicity. Copyright (C) 2009 John Wiley & Sons, Ltd.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据