Polyethylenimine (PEI)/siRNA-Mediated Gene Knockdown In Vitro and In Vivo
Polyethylenimine (PEI)/siRNA-Mediated Gene Knockdown In Vitro and In Vivo
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DOI：
10.1007/978-1-60761-588-0_18
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发表时间：
2010-01-01
期刊：
影响因子：
--
通讯作者：
Aigner, Achim
中科院分区：
文献类型：
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作者：
Hoebel, Sabrina;Aigner, Achim
自10年前发现以来，RNA干扰（RNAi）已成为敲除任何目标基因的标准方法。它由小干扰RNA（siRNA）介导，其触发mRNA降解的催化机制。因此，完整siRNA的递送对于RNAi的诱导至关重要。由于siRNA的物理化学和生物学性质，导致高不稳定性和差的细胞摄取，siRNA修饰和药物制剂已用于增强RNAi功效。这与SiRNA的体内递送特别相关，这仍然是RNAi实验或治疗应用的主要障碍。聚乙烯亚胺（PEI）是长度可变的水溶性、线性或支化合成聚合物，每三个位置上都有可质子化的氨基。我们已经表明，某些PEI能够与siRNA形成非共价复合物，介导其对溶核降解的保护以及增强其细胞摄取和细胞内释放。在本章中，PEI/siRNA复合物的制备和使用的各种体外和体内应用进行了描述。并给出了进行基因打靶实验和敲除效果分析的实例。
Since its discovery about 10 years ago, RNA interference (RNAi) has become an almost standard method for the knockdown of any target gene of interest. It is mediated by small interfering RNAs (siRNAs), which trigger a catalytic mechanism for mRNA degradation. Consequently, the delivery of intact siRNA is of critical importance for the induction of RNAi. Due to the physicochemical and biological properties of siRNAs, resulting in high instability and poor cellular uptake, siRNA modifications and pharmaceutical formulations have been used to enhance RNAi efficacy. This is particularly relevant for the in vivo delivery of siRNAs, which still poses a major hurdle for the experimental or therapeutic application of RNAi.Polyethylenimines (PEIs) are water-soluble, linear, or branched synthetic polymers of variable length with protonable amino groups in every third position. We have shown that certain PEIs are able to form noncovalent complexes with siRNAs, which mediate their protection against nucleolytic degradation as well as enhance their cellular uptake and intracellular release. In this chapter, the preparation and use of PEI/siRNA complexes for various in vitro and in vivo applications are described. Examples for conducting gene targeting experiments and the analysis of knockdown efficacies are given.