Methods and compositions for use in spliceosome mediated RNA trans-splicing

A recombinant expression vector wherein said vector expresses a nucleotide sequence comprising:

• a) two or more target binding domains that target binding of the pre-trans-splicing molecule to a target pre-mRNA;
• b) a 3′ splice region comprising a branchpoint, a pyrimidine tract and a 3′ splice acceptor site and a 5′ splice donor site;
• c) spacer regions that separate the 3′ splice region and the 5′ splice donor site from the target binding domains;
• and
• d) a nucleotide sequence to be trans-spliced to the target pre-mRNA;
• wherein said nucleic acid molecule is recognized by nuclear splicing components within the cell.

A cell comprising a nucleic acid molecule wherein said nucleic acid molecule comprises:

• a) two or more target binding domains that target binding of the pre-trans-splicing molecule to a target pre-mRNA;
• b) a 3′ splice region comprising a branch point, a pyrimidine tract and a 3′ splice acceptor site and a 5′ splice donor site;
• c) spacer regions that separate the 3′ splice region and the 5′ splice donor site from the target binding domains;
• and
• d) a nucleotide sequence to be trans-spliced to the target pre-mRNA;
• wherein said nucleic acid molecules is recognized by nuclear splicing components within the cell.

A cell comprising a recombinant expression vector wherein said vector expresses a nucleotide sequence comprising:

• a) two or more target binding domains that target binding of the pre-trans-splicing molecule to a target pre-mRNA;
• b) a 3′ splice region comprising a branchpoint, a pyrimidine tract and a 3′ splice acceptor site and a 5′ splice donor site;
• c) spacer regions that separate the 3′ splice region and the 5′ splice donor site from the target binding domains;
• and
• d) a nucleotide sequence to be trans-spliced to the target pre-mRNA;
• wherein said nucleic acid molecules is recognized by nuclear splicing components within the cell.

A nucleic acid molecule comprising:

• a) one or more target binding domains that target binding of the pre-trans-splicing molecule to a target pre-mRNA;
• b) a 5′ splice donor site;
• C) a spacer region that separates the 5′ splice donor site from the target binding domain;
• and
• d) a nucleotide sequence comprising the 5′ end of a gene to be trans-spliced to the target pre-mRNA;
• wherein said nucleic acid molecules is recognized by nuclear splicing components within the cell.

A recombinant expression vector wherein said vector expresses a nucleotide sequence comprising:

• a) one or more target binding domains that target binding of the pre-trans-splicing molecule to a target pre-mRNA;
• b) a 5′ splice donor site;
• c) a spacer region that separates the 5′ splice donor site from the target binding domain;
• and
• d) a nucleotide sequence comprising the 5′ end of a gene to be trans-spliced to the target pre-mRNA;
• wherein said nucleic acid molecules is recognized by nuclear splicing components within the cell.

A cell comprising a nucleic acid molecule wherein said nucleic acid molecule comprises:

• a) one or more target binding domains that target binding of the pre-trans-splicing molecule to a target pre-mRNA;
• b) a 5′ splice donor site;
• c) a spacer region that separates the 5′ splice donor site from the target binding domain;
• and
• d) a nucleotide sequence comprising the 5′ end of a gene to be trans-spliced to the target pre-mRNA;
• wherein said nucleic acid molecules is recognized by nuclear splicing components within the cell.

A cell comprising a recombinant expression vector wherein said vector expresses a nucleotide sequence comprising:

• a) one or more target binding domains that target binding of the pre-trans-splicing molecule to a target pre-mRNA;
• b) a 5′ splice donor site;
• c) a spacer region that separates the 5′ splice donor site from the target binding domain;
• and
• d) a nucleotide sequence comprising the 5′ end of a gene to be trans-spliced to the target pre-mRNA;
• wherein said nucleic acid molecules is recognized by nuclear splicing components within the cell.