Development of a specific siRNA delivery system into HeLa cells using an IgG-binding fusion protein
Development of a specific siRNA delivery system into HeLa cells using an IgG-binding fusion protein
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使用 IgG 结合融合蛋白开发 HeLa 细胞中的特异性 siRNA 递送系统
DOI：
10.1007/s10529-013-1299-y
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发表时间：
2013
期刊：
影响因子：
--
通讯作者：
Kobatake E
中科院分区：
文献类型：
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作者：
Bae J;Mie M;Kobatake E
需要稳定的载体来将siRNA递送至细胞。使用聚乙烯亚胺（PEI）作为基因载体已被广泛研究;然而，它不能提供足够的保护免受RNA酶降解，并且不适合靶向siRNA递送至特定细胞。在这项研究中，两个重复的Fc结合结构域的蛋白G（C2）被用来引入特异性抗体的PEI为基础的载体的siRNA。此外，我们使用了可以与siRNA结合的双链RNA结合结构域（DRBD）。该复合物由PEI、siRNA和构建的包含C2和DRBD结构域的融合蛋白TrxC 2DRBD组成，可保护siRNA免受RNase降解。此外，细胞特异性siRNA递送到HeLa细胞中可以通过与特异性抗体经由C2结构域的复合物融合来进行。
Stable carriers are required for delivering siRNA to cells. The use of polyethyleneimine (PEI) as gene carrier has been researched extensively; however, it does not provide sufficient protection from RNase degradation and is not suitable for targeted siRNA delivery to specific cells. In this study, two repeats of Fc binding domain of protein G (C2) were used to introduce a specific antibody to PEI-based carrier of siRNA. In addition, we used the double-stranded RNA binding domain (DRBD) that can bind to siRNA. The complex, consisting of PEI, siRNA and constructed fusion protein, TrxC2DRBD including C2 and DRBD domains, could protect siRNA from RNase degradation. Furthermore, cell specific siRNA delivery into HeLa cells could be performed by the complex fusion with specific antibodies via C2 domain.