Modified factor IX, and compositions, methods and uses for gene transfer to cells, organs, and tissues
AssigneeThe Children's Hospital of Philadelphia
InventorsKatherine A. High, Xavier Anguela
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Abstract
The invention relates to modified Factor IX coding sequence, expression cassette, vectors such as viral (e.g., lenti- or adeno-associated viral) vectors, and gene transfer methods and uses. In particular, to target Factor IX nucleic acid to cells, tissues or organs for expression (transcription) of Factor IX.
Claims65
A recombinant AAV (rAAV) vector comprising a capsid and a genome, wherein said capsid comprises a VP1 protein comprising the amino acid sequence of SEQ ID NO:4, and wherein said genome comprises a nucleic acid comprising a sequence encoding human Factor IX (FIX) protein, wherein the coding sequence of said nucleic acid is not naturally occurring, is at least 90% identical to SEQ ID NO:10, encodes the same human Factor IX protein encoded by SEQ ID NO:10, has a reduced number of CpG di-nucleotides compared to a wild-type sequence encoding human Factor IX protein and optionally is interrupted by an intron.
A recombinant AAV (rAAV) vector comprising a capsid and a genome, wherein said capsid comprises a VP1 protein comprising the amino acid sequence of SEQ ID NO:4, and wherein said genome comprises a nucleic acid comprising a sequence encoding human FIX protein, wherein the coding sequence of said nucleic acid is not naturally occurring, is at least 90% identical to SEQ ID NO:10, encodes the same human FIX protein encoded by SEQ ID NO:10, and optionally is interrupted by an intron.
A recombinant AAV (rAAV) vector for treating hemophilia B, comprising a capsid and a genome, wherein said capsid comprises a VP1 protein having the amino acid sequence of SEQ ID NO:4;
- and wherein said genome is single-stranded and comprises the following elements in 5′ to 3′ order:
- (a) a first AAV2 ITR,
- (b) an ApoE HCR-1 enhancer,
- (c) an AAT promoter,
- (d) a nucleic acid comprising a sequence encoding human FIX Padua variant, wherein the coding sequence of said nucleic acid is not naturally occurring, is at least 90% identical to SEQ ID NO:10, encodes the same human FIX protein encoded by SEQ ID NO:10, and optionally is interrupted by an intron,
- (e) a polyadenylation sequence;
- and
- (f) a second AAV2 ITR.
A packaging cell comprising (i) a nucleic acid comprising a sequence encoding human FIX protein, wherein the coding sequence of said nucleic acid is not naturally occurring, is at least 90% identical to SEQ ID NO:10, encodes the same human FIX protein encoded by SEQ ID NO:10, and optionally is interrupted by an intron, (ii) AAV rep protein, (iii) AAV capsid protein comprising the amino acid sequence of SEQ ID NO:4, and (iv) adenovirus helper protein(s).
A recombinant AAV (rAAV) vector comprising a capsid and a genome, wherein said capsid comprises a VP1 protein comprising the amino acid sequence of SEQ ID NO:4, and wherein said genome comprises at least one AAV ITR and a nucleic acid comprising a sequence encoding human FIX protein, wherein the coding sequence of said nucleic acid is not naturally occurring, is at least 90% identical to SEQ ID NO:10, encodes the same human FIX protein encoded by SEQ ID NO:10, and optionally is interrupted by an intron.
A recombinant AAV (rAAV) vector comprising a capsid and a genome, wherein said capsid comprises a VP1 protein comprising the amino acid sequence of SEQ ID NO:4, and wherein said genome comprises an AAV2 ITR, a liver-specific enhancer and promoter in operable linkage with a nucleic acid comprising a sequence encoding human FIX protein, wherein the coding sequence of said nucleic acid is not naturally occurring, is at least 90% identical to SEQ ID NO:10, encodes the same human FIX protein encoded by SEQ ID NO:10, and optionally is interrupted by an intron, a transcription terminator, and optionally a second AAV2 ITR.
A recombinant AAV (rAAV) vector for treating hemophilia B, comprising a capsid and a genome, wherein said capsid comprises a VP1 protein having the amino acid sequence of SEQ ID NO:4;
- and wherein said genome is comprises the following elements:
- (a) a first AAV2 ITR,
- (b) a liver specific enhancer and promoter,
- (c) a nucleic acid sequence encoding human FIX Padua variant, wherein the coding sequence of said nucleic acid is not naturally occurring, is at least 90% identical to SEQ ID NO:10, encodes the same human FIX protein encoded by SEQ ID NO:10, and is interrupted by an intron,
- (d) a polyadenylation sequence;
- and
- (e) a second AAV2 ITR.
Dependencies are inferred from the claim text.