University of Florida Research Foundation Inc

The State University of New York

Other

The University of North Carolina at Chapel Hill

The Uab Research Foundation (the University of Alabama at Birmingham)

Guy, John

Nicholas Muzyczka

Sergei Zolotukhin

William W Hauswirth

Tiehua Ni

Xiaohuai Zhou

Barry John Byrne

Richard Jude Samulski

Hector Ruben Mendez-Gomez

Sanford Leon Boye

Shannon Elizabeth Boye

Edgardo Rodriguez

Richard O Snyder

Edwin M Meyer

Damien Marsic

Terence R Flotte

Paul D Gamlin

John W Guy

Mark R Potter

Craig Allen Meyers

Kenneth I Berns

Irine Zolotukhin

Scott Loiler

Kenneth H Warrington

Vince A Chiodo

Corinna Burger

Kevin R Nash

Shaun R Opie

Thomas J Fraites

Denise Gavin

Corinne Abernathy

Daniel Pereira

Ronald Klein

Mike King

Paul L Hermonat

Yoshihisa Sakai

John Guy

Adeno associated viral (AAV) particles are emerging as a useful vehicle for gene delivery to various organs and tissues, one of them being the retina. Provided here are variant AAV (e.g., variant serotype 2 (AAV2)) capsid proteins and variant capsid protein containing particles with enhanced ability to transduce retinal cells.

Modified AAV capsid proteins and uses thereof

Provided herein are compositions and methods related to targeted delivery of a therapeutic or diagnostic agent to a subject utilizing an engineered receptor-ligand system, such as an engineered dockerin-cohesin system. As described herein, previously-developed targeted delivery systems for delivering therapeutic and diagnostic agents to a tissue of interest have drawbacks that have not been addressed to date. For example, with respect to the blood-brain barrier (BBB) and the blood-cerebrospinal fluid barrier (BCSFB), both of which hamper delivery of agents to the brain, others have relied on the use of endogenously expressed receptors, like the transferrin receptor, to assist the agent across the barriers.

Engineered receptor/ligand system for delivery of therapeutic agents

Disclosed are improved recombinant adeno-associated viral (rAAV) vectors having mutations in one or more capsid proteins. Exemplary vectors are provided that have altered affinity for heparin or heparin sulfate, as well as vectors, expression systems, and rAAV virions that lack functional VP2 protein expression, but are nevertheless, fully virulent. Also provided by the invention are rAAV vector-based compositions, virus particles, host cells, and pharmaceutical formulations that comprise them useful in the expression of selected therapeutic proteins, polypeptides, peptides, antisense oligonucleotides and/or ribozymes in selected mammals, including organs, tissues, and human host cells.

Raav expression systems for genetic modification of specific capsid proteins

Nucleic acid mitochondrial delivery systems are provided. These systems are important for delivery of genetic information to a mitochondrion and for treatment of mitochondrial related diseases.

Mitochondrial nucleic acid delivery systems

This invention provides a system for modeling neurodegenerative and other diseases through somatic gene transfer. In addition, methods of multiple gene transfer, disease analysis and drug testing are provided for.

Human disease modeling using somatic gene transfer

Vectors that encode Adeno-Associated Virus (AAV) Rep and Cap proteins of different serotypes and Adenovirus transcription products that provide helper functions were used to produce pseudotyped recombinant AAV (rAAV) virions. Purification methods generated pseudotyped rAAV virion stocks that were 99% pure with titers of 1&#xd7;10&#x2013;1&#xd7;10vector genomes/ml.

Production of pseudotyped recombinant AAV virions

Disclosed are methods for the isolation and purification of high-titer recombinant adeno-associated virus (rAAV) compositions. Also disclosed are methods for reducing or eliminating the concentration of helper adenovirus in rAAV samples. Methods are disclosed that provide highly-purified rAAV stocks having titers up to about 10 particles/ml at particle-to-infectivity ratios of less than 100 in processes that are accomplished about 24 hours or less.

Method of preparing recombinant adeno-associated virus compositions

The present invention provides temperature-sensitive (ts) adeno-associated virus (AAV) Rep78 and Rep68 proteins. In preferred embodiments, the ts AAV Rep78 and Rep68 proteins have missense mutations at amino acid positions 40, 42 and 44 that confer a temperature-sensitive phenotype. Also provided are nucleotide sequences and vectors encoding the inventive ts Rep proteins. In preferred embodiments, a hybrid adenovirus vector is provided that stably comprises a nucleotide sequence encoding a ts AAV Rep protein according to the invention. The present invention also provides methods of packaging AAV vectors and methods of ex vivo gene delivery using the ts Rep proteins of the invention. Further provided are cells containing the ts AAV Rep proteins, preferably stably integrated into the genome of the cell.

Temperature-sensitive regulation of viral vector production

Disclosed are methods for the isolation and purification of high-titer recombinant adeno-associated virus (rAAV) compositions. Also disclosed are methods for reducing or eliminating the concentration of helper adenovirus in rAAV samples. Methods are disclosed that provide highly-purified rAAV stocks having titers up to about 10.sup.13 particles/ml at particle-to-infectivity ratios of less than 100 in processes that are accomplished about 24 hours or less.

Disclosed are synthetic and 'humanized' versions of green fluorescent protein (GFP) genes adapted for high level expression in mammalian cells, especially those of human origin. Base substitutions are made in various codons in order to change the codon usage to one more appropriate for expression in mammalian cells. Recombinant vectors carrying such humanized genes are also disclosed. In addition, various methods for using the efficient expression of humanized GFP in mammalian cells and in animals are described.

Humanized green fluorescent protein genes and methods

A method for in vitro packaging of adeno-associated viral particles is described. The procedure involves the preparation of cell-free extracts containing all the essential components for packaging. Homogeneous purified substrate DNA for packaging may be prepared separately. The in vitro packaged AAV particles are useful in transduction of mammalian cells and for gene therapy in animals. In one described method, the DNA packaged into AAV particles is not limited by the size constraints characteristic of in vivo packaged AAV particles.

In vitro packaging of adeno-associated virus DNA

Growing community of inventors

Gainesville, FL, United States of America

Nicholas Muzyczka