Induction of exon skipping in eukaryotic cells

Abstract

Described is a method for at least in part decreasing the production of an aberrant protein in a cell, the cell comprising pre-mRNA comprising exons coding for the protein, by inducing so-called exon skipping in the cell. Exon-skipping results in mature mRNA that does not contain the skipped exon, which leads to an altered product of the exon codes for amino acids. Exon skipping is performed by providing a cell with an agent capable of specifically inhibiting an exon inclusion signal, for instance, an exon recognition sequence, of the exon. The exon inclusion signal can be interfered with by a nucleic acid comprising complementarity to a part of the exon. The nucleic acid, which is also herewith provided, can be used for the preparation of a medicament, for instance, for the treatment of an inherited disease.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS[0001]This application is a divisional of co-pending U.S. patent application Ser. No. 10 / 395,031, filed Mar. 21, 2003, now U.S. Pat. No. ______, which is a continuation of International Application PCT / NL01 / 00697, filed Sep. 21, 2001, designating the United States, published in English Mar. 28, 2002, as WO 02 / 024906 A1 and subsequently published with corrections Jan. 23, 2003, as WO 02 / 024906 C2, the contents of the entirety of each of which are hereby incorporated herein by this reference.TECHNICAL FIELD[0002]The invention relates to the fields of biotechnology and gene therapy.BACKGROUND[0003]Given the rapid advances of human genome research, professionals and the public expect that the near future will bring us, in addition to understanding of disease mechanisms and refined and reliable diagnostics, therapies for many devastating genetic diseases.[0004]While it is hoped that for some (e.g., metabolic) diseases, the improved insights will bri...

AI Technical Summary

Benefits of technology

[0018]Methods of the invention can be used in many ways. In one embodiment, a method described herein is used to at least in part decrease the production of an aberrant protein. Such proteins can, for instance, be oncoproteins or viral proteins. In many tumors, not only the presence of an oncoprotein but also its relative level of expression has been associated with the phenotype of the tumor cell. Similarly, not only the presence of viral proteins but also the amount of viral protein in a cell determines the virulence of a particular virus. Moreover, for efficient multiplication and spread of a virus, the timing of expression in the life cycle and the balance in the amount of certain viral proteins in a cell determines whether viruses are efficiently or inefficiently produced. Using a method described herein, it is possible to lower the amount of aberrant protein in a cell such that, for instance, a tumor cell becomes less tumorigenic (metastatic) and / or a virus-infected cell produces less virus.

[0019]In certain embodiments, a method described herein is used to modify the aberrant protein into a functional protein. In one embodiment, the functional protein is capable of performing a function of a protein normally present in a cell but absent in the cells to be treated. Very often, even partial restoration of function results in significantly improved performance of the cell thus treated. Due to the better performance, such cells can also have a selective advantage over unmodified cells, thus aiding the efficacy of the treatment.

Problems solved by technology

Hybridization of the oligos to other splice sites than the sites of the exon to be skipped of course could easily interfere with the accuracy of the splicing process.

On the other hand, the accuracy could be lacking due to the fact that two oligos (for the 5′ and the 3′ splice site) need to be used.

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