title: Promoter choice affects the potency of HIV-1 specific RNA interference.
abstract: RNA interference (RNAi) is mediated by small interfering (si) RNAs that target and degrade mRNA in a sequence-specific manner. Cellular expression of siRNA can be achieved by the use of expression cassettes driven by RNA polymerase III (pol III) promoters. Here, we demonstrate that a modified tRNA(met)-derived (MTD) promoter effectively drives the cellular expression of HIV-1-specific siRNA. We observed up to 56% greater inhibition of virus production when the MTD promoter was used to drive the expression of short hairpin (sh) RNA targeting the HIV-1 transactivator protein tat compared to cassettes containing other pol III promoters such as H1, U6+1 and U6+27. We conclude that the MTD promoter is ideally suited to drive intracellular expression of HIV-1 specific siRNA and may serve as an important component of future RNAi vector delivery systems.
authors.affiliation: Laboratory of Retrovirology, Division of Infectious Diseases, Department of Medicine, Brown Medical School, 4th floor, 55 Claverick Street, Providence, RI 02903, USA.
authors.forename: Daniel
authors.initials: D
authors.lastname: Boden
authors.forename: Oliver
authors.initials: O
authors.lastname: Pusch
authors.forename: Fredrick
authors.initials: F
authors.lastname: Lee
authors.forename: Lynne
authors.initials: L
authors.lastname: Tucker
authors.forename: Peter R
authors.initials: PR
authors.lastname: Shank
authors.forename: Bharat
authors.initials: B
authors.lastname: Ramratnam
mesh_terms: D002460:Cell Line; D015967:Gene Expression Regulation, Viral; D015696:Gene Products, tat / Q000235:genetics; D015497:HIV-1 / Q000235:genetics*; D006801:Humans; D011401:Promoter Regions, Genetic / Q000235:genetics*; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics / Q000378:metabolism; D012358:RNA, Transfer, Met / Q000235:genetics; D012367:RNA, Viral / Q000235:genetics / Q000378:metabolism; D014162:Transfection; D054322:tat Gene Products, Human Immunodeficiency Virus
chemical_list: D015696:Gene Products, tat; D034741:RNA, Small Interfering; D012358:RNA, Transfer, Met; D012367:RNA, Viral; D054322:tat Gene Products, Human Immunodeficiency Virus