STABLE ALTERATION OF PRE-MRNA SPLICING PATTERNS BY MODIFIED RNAS

摘要

The present invention provides a method of upregulating expression of a protein of interest (e.g., a native protein) in a cell, the cell containing a DNA encoding the protein, which DNA contains a mutation that causes downregulation of the protein by aberrant splicing in a pre-mRNA, wherein the DNA encodes the pre-mRNA; wherein the pre-mRNA contains a native intron having a first set of splice elements, which native intron is removed by splicing when the mutation is absent to produce a first mRNA encoding the protein; and wherein the pre-mRNA further contains an aberrant intron different from the native intron having a second set of splice elements, which aberrant intron is removed by splicing when the mutation is present to produce an aberrant second mRNA different from the first mRNA. The method comprises administering to the cell a gene transfer vector a heterologous oligonucleotide in the cell, the heterologous oligonucleotide comprising a nuclear localization element joined to an antisense oligonucleotide, which antisense oligonucleotide hybridizes to the pre-mRNA in the nucleus of the cell to create a duplex thereof under conditions which permit splicing, and wherein the antisense oligonucleotide blocks a member of the aberrant second set of splice elements so that the native intron is removed by splicing and the protein of interest is produced. Vectors and oligonucleotides useful for carrying out the method are also disclosed.