Means and methods for modulating NOTCH3 protein expression and/or the coding region of NOTCH3; compositions and use thereof in the treatment of CADASIL

摘要

The invention among other provides means and methods for modulating NOTCH3 expression and/or protein coding domain. In one aspect the invention provides a method for at least reducing an elevated level of NOTCH3 protein in a NOTCH3 expressing cell or the immediate vicinity thereof said method comprising providing said cell with an anti-sense oligonucleotide specific for NOTCH3 m RNA or pre-m RNA thereby decreasing production of said NOTCH3 protein or thereby altering the protein coding region in said NOTCH3 m RNA or pre-m RNA.

主权项

1. A method for producing an mRNA encoding a NOTCH3 protein having EGFL repeats with even numbers of cysteine residues from a pre-mRNA encoding a mutant NOTCH3 protein having an EGFL repeat with an uneven number of cysteine residues, comprising, providing a cell expressing said pre-mRNA with anti-sense oligonucleotides that are complementary to and hybridize with said pre-mRNA and wherein the anti-sense oligonucleotides are exon 4 and exon 5 internal oligonucleotides or directed toward the splice sites flanking the exons, and thereby inducing the exclusion of exon 4 and exon 5 from mRNA produced from said pre-mRNA, resulting in the cell producing an mRNA encoding a NOTCH3 protein having EGFL repeats with even numbers of cysteine residues;
wherein the produced mRNA comprises an exon 3-6 joining that is in frame and encodes a NOTCH3 protein in which the EGFL repeats all have an even number of cysteine residues, and wherein the oligonucleotides comprise a chemical modification that renders a duplex formed by the antisense oligonucleotides and the target pre-mRNA resistant to RNAseH.