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Production and discovery of novel recombinant adeno-associated viral vectors

dc.contributor.authorMueller, Christian
dc.contributor.authorZhong, Li
dc.contributor.authorRatner, Dmitry
dc.contributor.authorSena-Esteves, Miguel
dc.contributor.authorGao, Guangping
dc.date2022-08-11T08:10:14.000
dc.date.accessioned2022-08-23T17:00:46Z
dc.date.available2022-08-23T17:00:46Z
dc.date.issued2012-08-01
dc.date.submitted2012-08-14
dc.identifier.citation<p>Curr Protoc Microbiol. 2012 Aug;Chapter 14:Unit14D.1. DOI: 10.1002/9780471729259.mc14d01s26</p>
dc.identifier.issn1934-8533
dc.identifier.doi10.1002/9780471729259.mc14d01s26
dc.identifier.pmid22875565
dc.identifier.urihttp://hdl.handle.net/20.500.14038/43861
dc.description.abstractIn this unit, we describe the detailed procedure for a three-plasmid transfection method for rAAV production, and discuss its advantages, limitations, and troubleshooting techniques. We further discuss the rAAV purification process using CsCl gradients, as well as subsequent quality control methods using SDS-PAGE and real-time PCR to assess vector purity, packaging efficiency, and viral titer. Finally, we elaborate on a PCR-based strategy that can be used to discover novel AAV capsid sequences from primate tissue, which can be used to develop newer-generation rAAVs with a greater diversity of tissue tropism for clinical gene therapy. Curr. Protoc. Microbiol. 26:14D.1.1-14D.1.21. © 2012 by John Wiley & Sons, Inc.
dc.language.isoen_US
dc.relation<a href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=22875565&dopt=Abstract">Link to article in PubMed</a>
dc.relation.urlhttp://dx.doi.org/10.1002/9780471729259.mc14d01s26
dc.subjectDependovirus
dc.subjectGenetic Vectors
dc.subjectGene Therapy
dc.subjectAllergy and Immunology
dc.subjectBiomedical Engineering and Bioengineering
dc.subjectGenetics and Genomics
dc.subjectMedical Genetics
dc.subjectPediatrics
dc.subjectRespiratory Tract Diseases
dc.titleProduction and discovery of novel recombinant adeno-associated viral vectors
dc.typeBook Chapter
dc.source.booktitleCurrent Protocols in Microbiology
dc.source.volumeChapter 14
dc.identifier.legacycoverpagehttps://escholarship.umassmed.edu/peds_pulmonary/72
dc.identifier.contextkey3212184
html.description.abstract<p>In this unit, we describe the detailed procedure for a three-plasmid transfection method for rAAV production, and discuss its advantages, limitations, and troubleshooting techniques. We further discuss the rAAV purification process using CsCl gradients, as well as subsequent quality control methods using SDS-PAGE and real-time PCR to assess vector purity, packaging efficiency, and viral titer. Finally, we elaborate on a PCR-based strategy that can be used to discover novel AAV capsid sequences from primate tissue, which can be used to develop newer-generation rAAVs with a greater diversity of tissue tropism for clinical gene therapy. Curr. Protoc. Microbiol. 26:14D.1.1-14D.1.21. © 2012 by John Wiley & Sons, Inc.</p>
dc.identifier.submissionpathpeds_pulmonary/72
dc.contributor.departmentDepartment of Microbiology and Physiology Systems
dc.contributor.departmentDepartment of Neurology
dc.contributor.departmentDepartment of Medicine
dc.contributor.departmentDepartment of Pediatrics
dc.contributor.departmentGene Therapy Center
dc.source.pagesUnit14D.1


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