AAV DUAL VECTOR SYSTEMS FOR GENE THERAPY

摘要

The subject invention concerns materials and methods for treating eye diseases, such as Usher syndrome I B (USHIB). One aspect of the invention concerns AAV based dual vector systems that provide for expression of full-length proteins whose coding sequence exceeds the polynucleotide packaging capacity of individual AAV vector. In one embodiment, a vector system of the invention comprises i) a first AAV vector polynucleotide comprising an inverted terminal repeat at each end of the polynucleotide, and a suitable promoter followed by a partial coding sequence that encodes an N -terminal part of a full-length polypeptide and ii) a second AAV vector polynucleotide comprising an inverted terminal repeat at each end of the polynucleotide, and a partial coding sequence that encodes a C-terminal part of a full-length polypeptide, optionally followed by a polyadenylation (pA) signal sequence. The coding sequence in the first and second AAV vectors comprises sequence that overlaps. In another embodiment, a vector system of the invention comprises i) a first AAV vector polynucleotide comprising an inverted terminal repeat at each end, a suitable promoter followed by a partial coding sequence that encodes an N-terminal part of a full-length polypeptide followed by a splice donor site and intron and ii) a second AAV vector polynucleotide comprising an inverted terminal repeat at each end, followed by an intron and a splice acceptor site for the intron, followed by a partial coding sequence that encodes a C-terminal part of a full-length polypeptide, optionally followed by a polyadenylation (pA) signal sequence. The intron sequence in the first and second AAV vectors comprises sequence that overlaps.