Means and methods for the specific inhibition of genes in cells and tissue of the cns and/or eye

Abstract

Described is a method for the specific modulation of the expression of target genes in cells and/or tissues of the CNS and/or eye, wherein a composition comprising one or more doubled stranded oligoribonucleotides (dsRNA) is introduced into the cell, tissue or organism outside the blood-brain or blood-retina barriers. Furthermore, a method for the identification and validation of the function of a gene is provided, wherein the method provides a test cell, test tissue or test organism, which allow information to be gained on the function of the target gene. In addition, compositions and kits are described useful for those methods. In particular, components and methods for the diagnostic use and/or therapy of disorders related to the CNS and/or eye are provided which are based on RNA interference.

1 - 47 . (canceled) 
     
     
         48 . A method of delivering one or more oligoribonucleotides across the blood-brain or the blood-retina barrier comprising introducing a composition comprising one or more double-stranded oligoribonucleotides (dsRNA) into a cell, tissue or organism outside the blood-brain or blood-retina barriers, wherein said dsRNA is trafficked across said blood-brain or blood-retina barrier. 
     
     
         49 . The method of  claim 48 , wherein said method results in the provision of a test cell, test tissue or test organism, which can be maintained under conditions allowing the degradation of the corresponding mRNA of one or more of target genes by RNA interference. 
     
     
         50 . The method of  claim 49  further comprising identifying or validating the function of a gene, further comprising comparing a resulting phenotype produced in the test cell, test tissue or test organism with that of a suitable control, wherein the function of the gene is identified or validated. 
     
     
         51 . The method of  claim 48 , wherein the introduced dsRNS inhibits expression of a target gene that is expressed behind the blood-brain or blood-retina barrier. 
     
     
         52 . The method of  claim 51 , wherein one or more of said target genes encode a cellular mRNA. 
     
     
         53 . The method of  claim 48 , wherein the cells, or tissues are cells, or tissues of the eye. 
     
     
         54 . The method of  claim 48 , wherein said cells or tissues are cells or tissues of the inner segment of the eye ball. 
     
     
         55 . The method of  claim 54 , wherein said cells are retinal cells. 
     
     
         56 . The method of  claim 55 , wherein said cells are cells of the retinal pigment epithelium (RPE) or neurosensory retina cells. 
     
     
         57 . The method of  claim 48 , wherein one or more of said target genes are predominantly expressed in said cell or tissue. 
     
     
         58 . The method of  claim 48 , wherein the expression of one or more of said target genes is specific for said cell or tissue. 
     
     
         59 . The method of  claim 48 , wherein said dsRNA molecules are between 21 and 23 nucleotides in length. 
     
     
         60 . The method of  claim 48 , wherein said dsRNA molecules contain a terminal 3′-hydroxyl group. 
     
     
         61 . The method of  claim 48 , wherein said dsRNA molecules are chemically synthesized. 
     
     
         62 . The method of  claim 48 , wherein said dsRNA molecules represent an analogue of naturally occurring RNA. 
     
     
         63 . The method of  claim 62 , wherein said dsRNA analogues differ from a corresponding naturally occurring RNA by addition, deletion, substitution or modification of one or more nucleotides. 
     
     
         64 . The method of  claim 48 , wherein said dsRNA molecules inhibit target genes by posttranscriptional silencing. 
     
     
         65 . The method of  claim 48 , wherein said dsRNA molecules are encoded by a vector. 
     
     
         66 . The method of  claim 65 , wherein the expression of said dsRNA is under control of a cell or tissue specific promoter. 
     
     
         67 . The method of  claim 48 , wherein the dsRNA molecules are bound to other molecules, combined with one or more suitable carriers, or any combination thereof. 
     
     
         68 . The method of  claim 67 , wherein the carrier is selected from a micellar structure and a viral coat protein. 
     
     
         69 . The method of  claim 67 , wherein the dsRNA is bound to molecules selected from the group consisting of cationic porphyrins, cationic polyamines, polymeric DNA-binding cations, fusogenic peptides, and any combination thereof. 
     
     
         70 . The method of  claim 67 , wherein the dsRNA molecules are delivered continuously to the target cells or target tissues over a defined period of time after application. 
     
     
         71 . The method of  claim 48 , wherein said composition is introduced by a method selected from the group consisting of iontophoresis, retrobulbar application, systemic application, topical application to the eye, or a combination of any thereof. 
     
     
         72 . The method of  claim 48 , wherein the cells, tissues or organism is a vertebrate. 
     
     
         73 . The method of  claim 48 , wherein the cells, tissues or organism is mammalian. 
     
     
         74 . The method of  claim 48 , wherein the cells, tissues or organism are human. 
     
     
         75 . The method of  claim 48 , wherein the dsRNA contains two symmetrical 3′ overhangs of two nucleotides in length. 
     
     
         76 . The method of  claim 75 , wherein the overhangs comprise 2′-deoxy-thymidine. 
     
     
         77 . The method of  claim 51 , wherein the inhibition of target gene expression treats a retinal disease. 
     
     
         78 . The method of  claim 51 , wherein the inhibition of target gene expression treats a degenerative retinal disease. 
     
     
         79 . The method of  claim 78 , wherein the degenerative retinal disease is selected from the group consisting of: primary detachment of the retina, retinoblastoma, retinal astrocytoma, angiomatosis retinae, Coats disease, Eales disease, retinopathia centralis serosa, ocular albinism, retinitis pigmentosa, retinitis punctata albescens, Usher's syndrome, Leber's congenital amaurosis, cone dystrophy, vitelliforme macular degeneration, juvenile retinoschisis, North Carolina macular dystrophy, Sorsby fundus-dystrophy, Doyne's honeycombs, retinal dystrophy, Morbus Stargardt, Wagner's vitreoretinal degeneration and age-dependent macular degeneration. 
     
     
         80 . The method of  claim 78 , wherein the degenerative retinal disease is age-dependent macular degeneration. 
     
     
         81 . The method of  claim 68 , wherein the micellar structure is a liposome. 
     
     
         82 . The method of  claim 68 , wherein the viral coat protein is derived from a virus selected from the group consisting of a cytomegalovirus, an adeno-associated virus and an adenovirus.