The United States of America As Represented by the Department of Health

Johns Hopkins University

Targeted Genetics Corporation

The United States of America, As Represented by the Secretary, Department of Health and Human Services

Rikki Solow

Barrie J Carter

Terence Flotte

Sandra Afione

Terence R Flotte

William B Guggino

Terrence Flotte

Barrier J Carter

Described herein are constructions of recombinant DNA comprising modified adeno-associated virus (AAV) DNA sequences capable of functioning as a eukaryotic expression vector for expressing foreign DNA sequences using a novel transcription promoter comprising the termini of AAV DNA. It is shown that expression of a test reporter gene can be obtained from this vector in mammalian cells. It is further shown that this combination of vector and promoter can be used to introduce and express a human gene and correct a genetic defect in human cells resulting from malfunction of the mutant endogenous gene. Further, the vector can be used to correct the genetic defect by expressing a modified version of the human gene consisting of a fusion of part of the said gene and a synthetic sequence contained in the vector.

Modified adeno-associated virus vector capable of expression from a

Amino-terminally truncated cystic fibrosis transmembrane conductance

Adeno-associated virus (AAV) vectors may have utility for gene therapy but heretofore a significant obstacle has been the inability to generate sufficient quantifies of such recombinant vectors in amounts that would be clinically useful for human gene therapy application. Stable, helper-free AAV packaging cell lines have been elusive, mainly due to the activities of Rep protein, which down-regulates its own expression and reverses cellular immortalization. This invention provides packaging systems and processes for packaging AAV vectors that efficiently circumvent these problems by replacing the AAV p5 promoter with a heterologous promoter and that allow for substantially increased packaging efficiency.

Generation of high titers of recombinant AAV vectors

Growing community of inventors

Gaithersburg, MD, United States of America

Rikki Solow