title: Toward a gene therapy for dominant disease: validation of an RNA interference-based mutation-independent approach.
abstract: The intragenic heterogeneity encountered in many dominant disease-causing genes represents a significant challenge with respect to development of economically viable therapeutics. For example, 25% of autosomal dominant retinitis pigmentosa is caused by over 100 different mutations within the gene encoding rhodopsin, each of which could require a unique gene therapy. We describe here an RNA interference (RNAi)-based mutation-independent approach, targeting as an example murine rhodopsin. Native transcripts are suppressed by a single RNAi molecular species, whereas transcripts from replacement genes engineered at degenerate third-codon wobble positions are resistant to suppression. We demonstrate suppression of murine rhodopsin transcript by up to 90% with full concomitant expression of replacement transcript and establish the validity of this approach in cell culture, retinal explants, and mouse liver in vivo.
authors.affiliation: Ocular Genetics Unit, Department of Genetics, Trinity College Dublin, Dublin 2, Ireland. skiang@tcd.ie
authors.forename: Anna-Sophia
authors.initials: AS
authors.lastname: Kiang
authors.forename: Arpad
authors.initials: A
authors.lastname: Palfi
authors.forename: Marius
authors.initials: M
authors.lastname: Ader
authors.forename: Paul F
authors.initials: PF
authors.lastname: Kenna
authors.forename: Sophia
authors.initials: S
authors.lastname: Millington-Ward
authors.forename: Gerry
authors.initials: G
authors.lastname: Clark
authors.forename: Avril
authors.initials: A
authors.lastname: Kennan
authors.forename: Mary
authors.initials: M
authors.lastname: O'reilly
authors.forename: Lawrence C T
authors.initials: LC
authors.lastname: Tam
authors.forename: Aileen
authors.initials: A
authors.lastname: Aherne
authors.forename: Niamh
authors.initials: N
authors.lastname: McNally
authors.forename: Pete
authors.initials: P
authors.lastname: Humphries
authors.forename: G Jane
authors.initials: GJ
authors.lastname: Farrar
mesh_terms: D000818:Animals; D019556:COS Cells; D002469:Cell Separation; D002478:Cells, Cultured; D002522:Chlorocebus aethiops; D018076:DNA, Complementary / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D018274:Electroporation; D005434:Flow Cytometry; D020868:Gene Silencing; D005799:Genes, Dominant; D015316:Genetic Therapy / Q000379:methods*; D008099:Liver / Q000378:metabolism; D051379:Mice; D008957:Models, Genetic; D009154:Mutation; D011312:Pressure; D012313:RNA / Q000378:metabolism; D034622:RNA Interference; D012333:RNA, Messenger / Q000378:metabolism; D034741:RNA, Small Interfering / Q000378:metabolism; D012160:Retina / Q000378:metabolism; D012174:Retinitis Pigmentosa / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012243:Rhodopsin / Q000378:metabolism; D013997:Time Factors; D014162:Transfection
chemical_list: D018076:DNA, Complementary; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D012313:RNA; D012243:Rhodopsin