title: Generation of genetic constructs that simultaneously express several shRNAs.
abstract: RNAi has potential as an antiviral gene therapy strategy. Cassette constructs simultaneously expressing several siRNAs could prove to be the most efficient technique in developing gene therapy approaches for highly mutable viruses such as HIV-1. Here we describe a rapid and cost-saving protocol to generate cassettes that simultaneously express three siRNAs for repression of HIV-1 and CCR5 transcripts. siRNA biological activity was tested in a non-viral system, and exhibited both efficiency and specificity. Our results suggest this protocol can be used to rapidly generate cassette constructs for antiviral gene therapy applications.
authors: Department of Genome Organization, Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Moscow, Russia. ovkretova@mail.ru
authors: Olga V
authors: OV
authors: Kretova
authors: Department of Genome Organization, Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Moscow, Russia.
authors: Ildar R
authors: IR
authors: Alembekov
authors: Department of Genome Organization, Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Moscow, Russia.
authors: Nickolai A
authors: NA
authors: Tchurikov
mesh_terms: D001483:Base Sequence; D002478:Cells, Cultured; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000235:genetics*; D015497:HIV-1 / Q000235:genetics; D006801:Humans; D008957:Models, Genetic; D008969:Molecular Sequence Data; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics*
chemical_list: D034741:RNA, Small Interfering
keywords: HIV-1; RNAi; multiple shRNA cassettes; non-viral test system