Recombinant adenovirus expressing adeno-associated virus cap and rep proteins supports production of high-titer recombinant adeno-associated virus
نویسندگان
چکیده
منابع مشابه
Production of high-titer recombinant adeno-associated virus vectors in the absence of helper adenovirus.
Recently, efficient and long-term in vivo gene transfer by recombinant adeno-associated virus type 2 (rAAV) vectors has been demonstrated in a variety of tissues. Further improvement in vector titer and purity will expedite this in vivo exploration and provide preclinical information required for use in human gene therapy. In an effort to obtain higher titers, we constructed a novel AAV helper ...
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Adeno-associated viral (AAV) vectors are used for in vivo gene transfer in a number of preclinical models of genetic diseases (including large-animal models) and are currently being tested in clinical trials for treatment of hemophilia B and cystic fibrosis. Protocols for production of AAV vectors in a helper virus-free system are available and are based on transient transfection of HEK-293 cel...
متن کاملHigh-titer, wild-type free recombinant adeno-associated virus vector production using intron-containing helper plasmids.
Recombinant adeno-associated virus (rAAV) is capable of directing long-term, high-level transgene expression without destructive cell-mediated immune responses. However, traditional packaging methods for rAAV vectors are generally inefficient and contaminated with replication-competent AAV (rcAAV) particles. Although wild-type AAV is not associated with any known human diseases, contaminating r...
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In this application note, we describe a novel method to rapidly measure the size and concentration of Adeno-Associated Virus (AAV) utilizing the Nanoparticle Tracking Analysis (NTA) technique and a gold labeling method. By labeling AAV capsids with small gold nanoparticles, we were able to increase the amount of light scattered by each capsid, enabling rapid NTA characterization to determine th...
متن کاملScalable generation of high-titer recombinant adeno-associated virus type 5 in insect cells.
We established a method for production of recombinant adeno-associated virus type 5 (rAAV5) in insect cells by use of baculovirus expression vectors. One baculovirus harbors a transgene between the inverted terminal repeat sequences of type 5, and the second expresses Rep78 and Rep52. Interestingly, the replacement of type 5 Rep52 with type 1 Rep52 generated four times more rAAV5 particles. We ...
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ژورنال
عنوان ژورنال: Gene Therapy
سال: 2001
ISSN: 0969-7128,1476-5462
DOI: 10.1038/sj.gt.3301454