Abstract
Ribozymes, catalytic RNA molecules that cleave a complementary mRNA sequence, have potential as therapeutics for dominantly inherited disease. Twelve percent of American patients with the blinding disease autosomal dominant retinitis pigmentosa (ADRP) carry a substitution of histidine for proline at codon 23 (P23H) in their rhodopsin gene1, resulting in photoreceptor cell death from the synthesis of the abnormal gene product. Ribozymes can discriminate and catalyze the in vitro destruction of P23H mutant mRNAs from a transgenic rat model of ADRP (ref. 2). Here, we demonstrate that in vivo expression of either a hammerhead or hairpin ribozyme in this rat model considerably slows the rate of photoreceptor degeneration for at least three months. Catalytically inactive control ribozymes had less effect on the retinal degeneration. Intracellular production of ribozymes in photoreceptors was achieved by transduction with a recombinant adeno-associated virus (rAAV) incorporating a rod opsin promoter. Ribozyme-directed cleavage of mutant mRNAs, therefore, may be an effective therapy for ADRP and also may be applicable to other inherited diseases.
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References
Rosenfeld, P.J. & Dryja, T.P. in Molecular Genetics of Ocular Disease Vol. 1 (ed. Wiggs, J.L.) 99–126 (Wiley-Liss, New York, 1995).
Drenser, K.A., Timmers, A.M., Hauswirth, W.W. & Lewin, A.S. Ribozyme-targeted destruction of RNAs associated with ADRP. Inv. Ophthalmol. Vis. Sci. 39, 681–689 (1998).
Birikh, K.R., Heaton, P.A. & Eckstein, F. The structure, function and application of the hammerhead ribozyme. Eur. J. Biochem. 245, 1–16 (1997).
Rossi, J.J. Therapeutic applications of catalytic antisense RNAs (ribozymes). Ciba. Found. Symp. 209, 195–204 (1997).
Lieber, A. & Kay, M.A. Adenovirus-mediated expression of ribozymes in mice. J. Virol. 70, 3153–3158 (1996).
L'Huillier, P. et al. Efficient and specific ribozyme-mediated reduction of bovine alpha-lactalbumin expression in double transgenic mice. Proc. Natl. Acad. Sci. USA 93, 6698–6703 (1996).
Flannery, J.G. et al. Efficient photoreceptor-targeted gene expression in vivo by recombinant adeno-associated virus. Proc. Natl. Acad. Sci. USA 94, 6916–6921 (1997).
Xiao, X., Li, J. & Samulski, R.J. Efficient long-term transfer into muscle tissue of immunocompetent mice by adeno-associated virus vector. J. Virol. 70, 8098–8108 (1996).
Fisher, K.J. et al. Recombinant adeno-associated virus for muscle directed gene therapy. Nature Med. 3, 306–312 (1997).
Steinberg, R.H. et al. Transgenic rat models of inherited retinal degeneration caused by mutant opsin genes. Invest. Ophthalmol. Vis. Sci. 37, S698 (1996).
Michon, J.J., Li, Z.L., Shioura, N., Anderson, R.J. & Tso, M.O. A comparative study of methods of photoreceptor morphometry. Invest. Ophthalmol. Vis. Sci. 32, 280–284 (1991).
Faktorovich, E.G. Steinberg, R.H., Yasumura, D., Matthes, M.T. & LaVail, M.M. Photoreceptor degeneration in inherited dystrophy delayed by the basic fibroblast growth factor. Nature 347, 83–86 (1990).
Wen, R. et al. Injury-induced upregulation of bFGF and CNTF mRNAS in the rat retina. J. Neurosci. 15, 7377–7385 (1995).
Hormes, R. et al. The subcellular localization and length of hammerhead ribozymes determine efficacy in human cells. Nucleic Acids. Res. 25, 769–775 (1997).
Miyoshi, H., Takahashi, M., Gage, F.H. & Verma, I.M. Stable and efficient gene transfer into retina using an HIV-based lentiviral vector. Proc. Natl. Acad. Sci. 94, 10319–10323 (1997).
Mohand-Said, S. et al. Photoreceptor transplants increase host cone survival in the retinal degeneration (rd) mouse. Ophthalmic Res. 29, 290–297 (1997).
Huang, P.C., Gaitan, A.E., Hao, Y., Petters, R.M. & Wong, F. Cellular interactions implicated in the mechanism of photoreceptor degeneration in transgenic mice expressing a mutant rhodopsin gene. Proc. Natl. Acad. Sci. USA 90, 8484–8488 (1993).
Zolotukhin, S., Potter, M., Hauswirth, W.W., Guy, J. & Muzyczka, N. A “humanized” green fluorescent protein cDNA adapted for high-level expression in mammalian cells. J. Virol. 70, 4646–4654 (1996).
Deslardin, L.E. & Hauswirth, W.W. Developmentally important elements within the bovine opsin upstream region. Invest. Ophthalmol. Vis. Sci. 37, 154–165 (1996).
Altschuler, M., Tritz, R. & Hampel, A. A method for generating transcripts with defined 5′ and 3′ ermini by autolytic processing. Gene 122, 85–90 (1992).
Bertrand, E. et al. The expression cassette determines the functional activity of ribozymes in mammalian cells by controlling their intracellular localization. RNA 3, 75–88 (1997).
LaVail, M.M. & Battelle, B.A. Influence of eye pigmentation and light deprivation on inherited retinal dystrophy in the rat. Exp. Eye Res. 21, 167–192 (1975).
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Lewin, A., Drenser, K., Hauswirth, W. et al. Ribozyme rescue of photoreceptor cells in a transgenic rat model of autosomal dominant retinitis pigmentosa. Nat Med 4, 967–971 (1998). https://doi.org/10.1038/nm0898-967
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DOI: https://doi.org/10.1038/nm0898-967
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