Abstract: The present invention provides methods and compositions for generating novel nucleic acid molecules through targeted spliceosomal mediated RNA trans-splicing. The compositions of the invention include pre-trans-splicing molecules (PTMs) designed to interact with a SERPINA1 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). In particular, the PTMs of the present invention include those genetically engineered to interact with SERPINA1 target pre-mRNA so as to result in correction of SERPINA1 genetic defects responsible for AAT deficiency. The PTMs of the invention may also comprise sequences that are processed out of the PTM to yield duplex siRNA molecules directed specifically to mutant SERPIN A1 mRNAs. Such duplexed siRNAs are designed to reduce the accumulation of toxic AAT protein in liver cells.

Abstract: The present invention provides methods and compositions for generating novel nucleic acid molecules through RNA trans-splicing that target a highly expressed pre-mRNA and contain the coding sequence for antibody polypeptide(s). The compositions of the invention include pre-trans-splicing molecules (PTMs) designed to interact with the target precursor messenger RNA molecule (target pre-mRNA) that is abundantly expressed or tumor specific and mediate a trans-splicing reaction resulting in the generation of novel chimeric RNA molecule (chimeric RNA) capable of encoding an antibody polypeptide. The invention provides for the in vivo production of chimeric RNA molecules that encode and result in the production of an antibody polypeptide that is therapeutically effective against, for example, infectious agents, cancer cells, transplantation antigens, rheumatoid arthritis, etc.

Abstract: Methods and compositions for generating novel nucleic acid molecules through targeted spliceosome mediated RNA trans-splicing that result in expression of a apoAI protein, an apoAI variant, the preferred embodiment referred to herein as the apoAI Milano variant, a pre-pro-apoAI or an analogue of apoAI. The methods and compositions include pre-trans-splicing molecules (PTMs) designed to interact with a 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) capable of encoding apoAI, the apoAI Milano variant, or an analogue of apoAI. The expression of this apoAI protein results in protection against vascular disorders resulting from plaque build up, i.e., atherosclerosis, strokes and heart attacks.

Abstract: The present invention provides methods and compositions for generating novel nucleic acid molecules through RNA trans-splicing that target a highly expressed pre-mRNA and contain the coding sequence for antibody polypeptide(s). The compositions of the invention include pre-trans-splicing molecules (PTMs) designed to interact with the target precursor messenger RNA molecule (target pre-mRNA) that is abundantly expressed or tumor specific and mediate a trans-splicing reaction resulting in the generation of novel chimeric RNA molecule (chimeric RNA) capable of encoding an antibody polypeptide. The invention provides for the in vivo production of chimeric RNA molecules that encode and result in the production of an antibody polypeptide that is therapeutically effective against, for example, infectious agents, cancer cells, transplantation antigens, rheumatoid arthritis, etc.

Abstract: The present invention provides methods and compositions for generating novel nucleic acid molecules through RNA trans-splicing that target a highly expressed pre-mRNA and contain the coding sequence of a protein or polypeptide of interest. The compositions of the invention include pre-trans-splicing molecules (PTMs) designed to interact with the target precursor messenger RNA molecule (target pre-mRNA) that is abundantly expressed, and mediate a trans-splicing reaction resulting in the generation of novel chimeric RNA molecule (chimeric RNA) capable of encoding a protein or polypeptide of interest. The invention provides for the in vivo production of chimeric RNA molecules that encode and result in the production of a protein or polypeptide of interest.

Abstract: The invention provides novel processes for manufacturing autologous T cells, transducing T cells and expanding the transduced T cell population.

Abstract: The present invention relates to methods and compositions for the efficient identification of one or more functionalities of a product encoded by a nucleic acid sequence of interest. The methods utilize the abilities to over and/or under express the product in a cell, as well as the combination of these results, to permit the identification of at least one of the product's cellular or in vivo functionality.

Abstract: This invention is directed to a soluble complex of ligands that binds to surface molecules of hemopoietic cells and result in their activation or expansion. The complex may be used in the activation and/or expansion of hemopoietic cells, optionally in combination with their transduction. The complex of ligands bind at least two cell surface molecules, such as one that plays a role in cell-cell adhesion and one that may or may not activate or stimulate the cell to promote growth and/or proliferation after binding to a ligand. A complex of ligands that bind two hemopoietic cell stimulatory molecules is also provided. The invention further provides for the use of the complex to target vectors to hemopoietic cells.

Abstract: The invention relates to a method of increasing vector transduction in target cells. The invention provides for the recombinant engineering of a packaging cell line to be capable of expressing one or more membrane proteins which facilitate binding to, and activation of, a target cell. The invention also provides for recombinant engineering of a cell that endogenously expresses one or more such membrane proteins into a packaging cell line. A vector packaged into viral particles via use of such cell lines would comprise an outer envelope containing these proteins. The particles would be specifically suited for binding and targeting to a target cell to facilitate transduction thereof with the vector. The target cell may also be simultaneously activated (stimulated) by the packaged vector in the absence of exogenously supplied stimulatory molecules.

Abstract: The present invention provides methods, as well as compositions related thereto, for the efficient transduction of cells using viral vectors. The efficiency of transduction is increased by contacting the cell to be transduced with one or more molecules that bind the cell surface. Contact with a cell surface binding molecule may occur before, after, or simultaneously with contact between the viral vector and the cell. The transduced vectors may be constructed to express a gene of interest, permitting the transduced cells to be used as therapeutic and prophylactic agents.