University of Pennsylvania

The University of North Carolina at Chapel Hill

Other

Neurologix, Inc.

Ivygen Corporation

Weidong Xiao

James M Wilson

Richard Jude Samulski

Joseph E Rabinowitz

Matthew J During

Wenjing Cao

Biao Dong

Lei Cao

In one aspect, present invention provides a recombinant mutant human factor VIII having increased expression and/or secretion as compared to wild-type factor VIII. In certain embodiments, the recombinant factor VIII includes one or more amino acid substitution(s) selected from the group consisting of I86, Y105, A108, D115, Q117, F129, G132, H134, M147 and L152. In other aspects, the present invention provides FVIII encoding nucleic acids, FVIII-expression vectors, as well as methods of using the modified FVIII genes in the treatment of FVIII deficiencies, such as hemophilia A.

Mutant factor VIII compositions and methods

The nucleic acid sequences of adeno-associated virus (AAV) serotype 1 are provided, as are vectors and host cells containing these sequences and functional fragments thereof. Also provided are methods of delivering genes via AAV-1 derived vectors.

Adeno-associated virus serotype I nucleic acid sequences, vectors and host cells containing same

A method for the production of a replication-deficient recombinant virus vector is disclosed. The replication-deficient recombinant virus vector has a recombinant virus genome with one or more defective viral genes. The method comprises infecting a host cell with a carrier virus having a carrier virus genome encoding one or more trans factors or variants thereof, incubating the infected host cell for a desired period of time, and isolating the replication-deficient recombinant virus vector. The carrier virus is a cytoplasmic virus that retains the carrier virus genome in the cytoplasm of the host cell. The host cell contains the recombinant viral genome and retains the recombinant viral genome in a nucleus of the host cell. Also disclosed is a carrier virus for the production of a replication-deficient recombinant virus vector.

Methods and compositions for the production of recombinant virus vectors

Adeno-associated virus serotype 1 nucleic acid sequences, vectors and host cells containing same

The present invention provides genetically-engineered parvovirus capsids and viruses designed to introduce a heterologous gene into a target cell. The parvoviruses of the invention provide a repertoire of vectors with altered antigenic properties, packaging capabilities, and/or cellular tropisms as compared with current AAV vectors.

Virus vectors and methods of making and administering the same

Methods for efficient production of recombinant AAV employ a host cell containing a first nucleic acid molecule comprising from 5' to 3&#x2032;, a parvovirus P5 promoter, a spacer, an AAV rep sequence and an AAV cap gene sequence, wherein said spacer is of sufficient size to reduce expression of the rep78 and rep68 gene products; a second nucleic acid molecule comprising a minigene comprising a transgene flanked by AAV inverse terminal repeats (ITRs) and under the control of regulatory sequences directing expression thereof in a host cell; and helper functions essential to the replication and packaging of rAAV, which functions are not provided by the first or second nucleic acid molecules. Host cells and molecule constructs are also described.

Methods and vector constructs useful for production of recombinant AAV

An improved tet-repressible system is described in which the transgene is expressed under the control of a promoter which is activated upon binding of a fusion protein composed of a reverse tet repressor/activation domain to tet operator sequences located immediately upstream of the transgene promoter. The tet operator sequences are substantially free of interferon inducible response elements (ISRE). The reverse tet repressor is fused to an activation domain which lacks signals for protein clearance, thus extending expression of the fusion protein. Suitably, the tetO sequences are immediately upstream of a quiet promoter.

Recombinant parvovirus vectors and method of making

Methods for efficient production of recombinant AAV employ a host cell containing a first nucleic acid molecule comprising from 5&prime; to 3&prime;, a parvovirus P5 promoter, a spacer, an AAV rep sequence and an AAV cap gene sequence, wherein said spacer is of sufficient size to reduce expression of the rep78 and rep68 gene products; a second nucleic acid molecule comprising a minigene comprising a transgene flanked by AAV inverse terminal repeats (ITRs) and under the control of regulatory sequences directing expression thereof in a host cell; and helper functions essential to the replication and packaging of rAAV, which functions are not provided by the first or second nucleic acid molecules. Host cells and molecule constructs are also described.

The present invention provides methods and compositions for preparations of recombinant parvovirus virions with a reduced number of replication competent particles. The compositions of the present invention include nucleic acids encoding parvovirus helper functions which contain at least one non-native intron sequence. The present invention also includes helper function vectors, host cells transfected with the helper function vectors, methods of using the helper function vectors, and recombinant parvovirus virions produced by such methods.

Growing community of inventors

Fort Washington, PA, United States of America

Weidong Xiao