title: Generation of hairpin-based RNAi vectors for biological and therapeutic application.
abstract: RNA interference (RNAi) is a natural process of gene silencing mediated by small RNAs. Shortly after the discovery of the RNAi mechanism, scientists devised various methods of delivering small interfering RNAs (siRNAs) capable of co-opting the endogenous RNAi machinery and suppressing target gene expression based on sequence complementarity. RNAi has since become a powerful tool to study gene function and is being investigated as a potential therapeutic approach to treat a vast array of human diseases (e.g., cancer, viral infections, and dominant genetic disorders). Among the available RNAi vectors are hairpin-based expression platforms (short-hairpin RNAs and artificial microRNAs) designed to mimic endogenously expressed inhibitory RNAs. These RNAi vectors are capable of achieving long-term potent gene silencing in vitro and in vivo. Here, we describe methods to design and generate these hairpin-based vectors and briefly review considerations for downstream applications.
authors.affiliation: Department of Internal Medicine, University of Iowa, Iowa City, Iowa, USA.
authors.forename: Ryan L
authors.initials: RL
authors.lastname: Boudreau
authors.forename: Beverly L
authors.initials: BL
authors.lastname: Davidson
mesh_terms: D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular / Q000379:methods; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors; D006801:Humans; D035683:MicroRNAs / Q000235:genetics / Q000302:isolation & purification; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics*; D014780:Viruses / Q000235:genetics
chemical_list: D035683:MicroRNAs; D034741:RNA, Small Interfering