Wang Ya-Hui, Hou Yu-Wun, Lee Han-Jung
Department of Life Science, National Dong Hwa University, No. 1, Sec. 2, Da-Hsueh Road, Shoufeng, Hualien 97401, Taiwan.
J Biochem Biophys Methods. 2007 Jun 10;70(4):579-86. doi: 10.1016/j.jbbm.2007.01.010. Epub 2007 Jan 30.
RNA interference has recently become a useful research tool for the studies of gene functions, regulations, and therapies. The double-stranded RNA is utilized to induce the sequence-specific gene silencing. To achieve this goal of specific gene silencing, a proper delivery system of siRNA is highly demanded. A number of approaches for delivering siRNA have been explored over the last few years. In the present study, we demonstrated a simple peptide-based siRNA delivery system in mammalian cells. A GC-EGFP cell line stably expressing enhanced green fluorescent protein was established from stable transfection of human gastric carcinoma cells. The synthetic nona-arginine peptide, an arginine-rich intracellular delivery peptide, or called protein transduction domain peptide, could noncovalently form stable complexes with EGFP siRNA and deliver these mixtures into cells. After entry, siRNA appeared to stay in perinuclear regions within cell, and ultimately fulfilled its targeted egfp gene silencing. These data were in consonance with that RNA-induced silencing complex components could be also localized to these perinuclear regions, creating a focal point for RNA interference factories. In the future, this non-toxic peptide may be proved to be a useful tool for the delivery of exogenous siRNA in RNA interference research.
RNA干扰最近已成为研究基因功能、调控和治疗的一种有用的研究工具。双链RNA被用于诱导序列特异性基因沉默。为实现这种特异性基因沉默的目标，对小干扰RNA（siRNA）合适的递送系统有很高的要求。在过去几年里，人们探索了多种递送siRNA的方法。在本研究中，我们展示了一种在哺乳动物细胞中基于肽的简单siRNA递送系统。通过稳定转染人胃癌细胞建立了稳定表达增强型绿色荧光蛋白的GC-EGFP细胞系。合成的九聚精氨酸肽，一种富含精氨酸的细胞内递送肽，或称蛋白质转导结构域肽，能够与EGFP siRNA非共价形成稳定复合物，并将这些混合物递送至细胞内。进入细胞后，siRNA似乎停留在细胞内核周区域，并最终实现其对egfp基因的靶向沉默。这些数据与RNA诱导沉默复合体组分也可定位于这些核周区域相符，从而为RNA干扰工厂创造了一个焦点。未来，这种无毒肽可能被证明是RNA干扰研究中递送外源性siRNA的一种有用工具。
