In vitro transfection using cyclodextrin-containing polycations

Numerous nonviral systems have been developed for the delivery of nucleic acids to cultured cells and to particular cell types in vivo. These systems vary with regard to their toxicity, immunogenicity, and ability to target particular cell surface receptors and/or cell types. A class of linear cationic polymers containing the sugar β-cyclodextrin has been shown to be effective at delivering a variety of nucleic acids in vivo, including plasmid DNA, DNAzymes, and short interfering RNAs (siRNAs). This protocol describes a procedure for the use of cyclodextrin-containing polycations in vitro. For in vitro transfection, salt stabilization and cell targeting are unnecessary, so polyplexes consist of cyclodextrin-containing polycations and nucleic acids only.