title: Promoter Targeting shRNA Suppresses HIV-1 Infection In vivo Through Transcriptional Gene Silencing.
abstract: Despite prolonged and intensive application, combined antiretroviral therapy cannot eradicate human immunodeficiency virus (HIV)-1 because it is harbored as a latent infection, surviving for long periods of time. Alternative approaches are required to overcome the limitations of current therapy. We have been developing a short interfering RNA (siRNA) gene silencing approach. Certain siRNAs targeting promoter regions of genes induce transcriptional gene silencing. We previously reported substantial transcriptional gene silencing of HIV-1 replication by an siRNA targeting the HIV-1 promoter in vitro. In this study, we show that this siRNA, expressed as a short hairpin RNA (shRNA) (shPromA-JRFL) delivered by lentiviral transduction of human peripheral blood mononuclear cells (PBMCs), which are then used to reconstitute NOJ mice, is able to inhibit HIV-1 replication in vivo, whereas a three-base mismatched variant (shPromA-M2) does not. In shPromA-JRFL-treated mice, HIV-1 RNA in serum is significantly reduced, and the ratio of CD4(+)/CD8(+) T cells is significantly elevated. Expression levels of the antisense RNA strand inversely correlates with HIV-1 RNA in serum. The silenced HIV-1 can be reactivated by T-cell activation in ex vivo cultures. HIV-1 suppression is not due to offtarget effects of shPromA-JRFL. These data provide "proof-of principle" that an shRNA targeting the HIV-1 promoter is able to suppress HIV-1 replication in vivo.Molecular Therapy-Nucleic Acids (2013) 2, e137; doi:10.1038/mtna.2013.64; published online 3 December 2013.
authors.affiliation: St. Vincent's Centre for Applied Medical Research, Darlinghurst, New South Wales, Australia.
authors.forename: Kazuo
authors.initials: K
authors.lastname: Suzuki
authors.forename: Shinichiro
authors.initials: S
authors.lastname: Hattori
authors.forename: Katherine
authors.initials: K
authors.lastname: Marks
authors.forename: Chantelle
authors.initials: C
authors.lastname: Ahlenstiel
authors.forename: Yosuke
authors.initials: Y
authors.lastname: Maeda
authors.forename: Takaomi
authors.initials: T
authors.lastname: Ishida
authors.forename: Michelle
authors.initials: M
authors.lastname: Millington
authors.forename: Maureen
authors.initials: M
authors.lastname: Boyd
authors.forename: Geoff
authors.initials: G
authors.lastname: Symonds
authors.forename: David A
authors.initials: DA
authors.lastname: Cooper
authors.forename: Seiji
authors.initials: S
authors.lastname: Okada
authors.forename: Anthony D
authors.initials: AD
authors.lastname: Kelleher