Methods and compositions for use in spliceosome mediated RNA trans-splicing
OwnerLloyd G. Mitchell, Mariano A. Garcia-Blanco +1inventors
InventorsLloyd G. Mitchell, Mariano A. Garcia-Blanco, Madaiah Puttaraju
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Abstract
The present invention provides methods and compositions for delivery of synthetic pre-trans-splicing molecules (synthetic PTMs) into a target cell. The compositions of the invention include synthetic pre-trans-splicing molecules (PTMs) with enhanced stability against chemical and enzymatic degradation. The synthetic PTMs are designed to interact with a natural target precursor messenger RNA molecule (target pre-mRNA) and mediate a trans-splicing reaction resulting in the generation of a novel chimeric RNA molecule (chimeric RNA).
Claims18
A method of producing a chimeric RNA molecule in a cell comprising:
- contacting a cell with a nucleic acid molecule recognized by nuclear splicing components wherein said nucleic acid molecule comprises:
- a) one or more target binding domains that target binding of the nucleic acid molecule to a pre-mRNA expressed within the cell;
- b) a 3′ splice region comprising a branch point, a pyrimidine tract and a 3′ splice acceptor site;
- and
- c) 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 method of producing a chimeric RNA molecule in a cell comprising:
- contacting a cell with a nucleic acid molecule recognized by nuclear splicing components wherein said nucleic acid molecule comprises:
- a) one or more target binding domains that target binding of the nucleic acid molecule to a pre-mRNA expressed within the cell;
- b) a 3′ splice acceptor site;
- and
- c) 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 method of producing a chimeric RNA molecule in a cell comprising contacting a cell with a nucleic acid molecule recognized by nuclear splicing components wherein said nucleic acid molecule comprises:
- a) one or more target binding domains that target binding of the nucleic acid molecule to a pre-mRNA expressed within the cell;
- b) a 5′ splice site;
- and
- c) 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 method of producing a chimeric RNA molecule in a cell comprising:
- contacting a cell with a nucleic acid molecule recognized by nuclear splicing components wherein said nucleic acid molecule comprises:
- a) one or more target binding domains that target binding of the nucleic acid molecule to a pre-mRNA expressed within the cell;
- b) a 5′ donor site;
- c) 3′ splice acceptor site;
- and
- c) 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.
Dependencies are inferred from the claim text.