Department of Environmental Chemistry and Engineering, Interdisciplinary Graduate School of Science and Engineering, Tokyo Institute of Technology, 4259 Nagatsuta, Midori-ku, Yokohama, 226-8502, Japan,
Biotechnol Lett. 2013 Dec;35(12):2081-9. doi: 10.1007/s10529-013-1299-y. Epub 2013 Aug 2.
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.
稳定载体是将 siRNA 递送到细胞所必需的。聚亚乙基亚胺（PEI）作为基因载体已经得到了广泛的研究；然而，它不能提供足够的保护以防止 RNA 酶的降解，也不适合将靶向 siRNA 递送到特定的细胞。在这项研究中，使用了蛋白 G（C2）的两个重复 Fc 结合域来引入针对 siRNA 的 PEI 载体的特异性抗体。此外，我们使用了双链 RNA 结合域（DRBD），它可以与 siRNA 结合。由 PEI、siRNA 和构建的融合蛋白 TrxC2DRBD（包括 C2 和 DRBD 结构域）组成的复合物可以保护 siRNA 免受 RNA 酶的降解。此外，通过 C2 结构域与特异性抗体的复合物融合，可以将特定的 siRNA 递送到 HeLa 细胞中。
