Polyethylenimine (PEI)/siRNA-mediated gene knockdown in vitro and in vivo.
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.
Höbel
Sabrina
S
Department of Pharmacology and Toxicology, School of Medicine, Philipps-University, Marburg, Germany.
Aigner
Achim
A
D000818:Animals; D055785:Gene Knockdown Techniques / Q000379:methods*; D006801:Humans; D051379:Mice; D009368:Neoplasm Transplantation; D011094:Polyethyleneimine / Q000378:metabolism*; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics*; D014183:Transplantation, Heterologous
D034741:RNA, Small Interfering; D011094:Polyethyleneimine