Inhibition of human immunodeficiency virus type 1 by RNA interference using long-hairpin RNA.
Inhibition of virus replication by means of RNA interference has been reported for several important human pathogens, including human immunodeficiency virus type 1 (HIV-1). RNA interference against these pathogens has been accomplished by introduction of virus-specific synthetic small interfering RNAs (siRNAs) or DNA constructs encoding short-hairpin RNAs (shRNAs). Their use as therapeutic antiviral against HIV-1 is limited, because of the emergence of viral escape mutants. In order to solve this durability problem, we tested DNA constructs encoding virus-specific long-hairpin RNAs (lhRNAs) for their ability to inhibit HIV-1 production. Expression of lhRNAs in mammalian cells may result in the synthesis of many siRNAs targeting different viral sequences, thus providing more potent inhibition and reducing the chance of viral escape. The lhRNA constructs were compared with in vitro diced double-stranded RNA and a DNA construct encoding an effective nef-specific shRNA for their ability to inhibit HIV-1 production in cells. Our results show that DNA constructs encoding virus-specific lhRNAs are capable of inhibiting HIV-1 production in a sequence-specific manner, without inducing the class I interferon genes.
Konstantinova
P
P
Department of Human Retrovirology, Academic Medical Center K3-110, University of Amsterdam, Meibergdreef 15, 1105 AZ Amsterdam, The Netherlands.
de Vries
W
W
Haasnoot
J
J
ter Brake
O
O
de Haan
P
P
Berkhout
B
B
D000818:Animals; D045744:Cell Line, Tumor; D002522:Chlorocebus aethiops; D015870:Gene Expression; D015693:Gene Products, nef / Q000235:genetics; D020868:Gene Silencing; D005818:Genetic Engineering; D015316:Genetic Therapy / Q000379:methods*; D015658:HIV Infections / Q000628:therapy*; D015497:HIV-1 / Q000235:genetics*; D006801:Humans; D007372:Interferons / Q000096:biosynthesis; D012313:RNA / Q000235:genetics*; D034622:RNA Interference; D016372:RNA, Antisense / Q000008:administration & dosage*; D034741:RNA, Small Interfering / Q000235:genetics; D012367:RNA, Viral / Q000235:genetics; D014162:Transfection / Q000379:methods; D014709:Vero Cells; D014779:Virus Replication / Q000235:genetics; D054311:nef Gene Products, Human Immunodeficiency Virus
D015693:Gene Products, nef; D016372:RNA, Antisense; D034741:RNA, Small Interfering; D012367:RNA, Viral; D054311:nef Gene Products, Human Immunodeficiency Virus; C514837:nef protein, Human immunodeficiency virus 1; D012313:RNA; D007372:Interferons