公开(公告)号：US6153436A

公开(公告)日：2000-11-28

申请号：US4877

申请日：1998-01-09

申请人： Paul L. Hermonat ,  J. Gerald Quirk ,  Brian Bishop ,  Han Li

发明人： Paul L. Hermonat ,  J. Gerald Quirk ,  Brian Bishop ,  Han Li

IPC分类号： A61K48/00 ,  C12N15/864 ,  C12N15/86

CPC分类号： C12N15/86 ,  A61K48/00 ,  C12N2750/14143

摘要： The present invention provides a method of human gene therapy using AAV vectors with essentially wildtype phenotype. Genes of 900 bases or less can be inserted into wildtype AAV and still allow the resulting vector to have a largely wildtype phenotype. For example, several antisense genes could be inserted and still allow packaging. Such wildtype vectors have several advantages. First, high titers of such vectors is easy to accomplish. Second, the vectors, since they include the Rep78 gene, integrate specifically into human chromosome 19. Third, such vectors, being wildtype, spread after their initial introduction. Another method for use of large wildtype AAV genomes is as complementors for fully defective AAV vectors. Such complementors can be delivered by virus infection and, be introduced easily into 100% of the cells used to produce virus. Viral infection is superior to synthetic techniques for introducing DNA into tissue culture producer cells. When large essentially wildtype AAV complementor virus are used in conjunction with AAV vector virus allowing for the introduction of both vectors into all cells, high titers of recombinant AAV virus is achieved.

摘要翻译： 本发明提供使用基本上野生型表型的AAV载体的人基因治疗方法。 将900个碱基以下的基因插入到野生型AAV中，并且仍然允许所得载体具有很大的野生型表型。 例如，可以插入几个反义基因并仍然允许包装。 这种野生型载体具有几个优点。 首先，这种载体的高滴度很容易完成。 第二，载体，因为它们包括Rep78基因，特异性整合入人染色体19。第三，这种载体，野生型，在其初次引入后扩散。 使用大型野生型AAV基因组的另一种方法是完全缺陷的AAV载体的补体。 这样的补体可以通过病毒感染递送，并且容易地引入用于产生病毒的100％的细胞中。 病毒感染优于将DNA引入组织培养生产细胞的合成技术。 当大的基本野生型AAV补体病毒与AAV载体病毒结合使用时，允许将两种载体引入所有细胞，实现了高效价的重组AAV病毒。