Site-specific integration of bacterial artificial chromosomes into human cells

Bradley McColl, Sara Howden, Jim Vadolas

Research output: Chapter in Book/Report/Conference proceedingChapter (Book)Otherpeer-review

Abstract

Gene therapy of inherited diseases requires long-term maintenance of the corrective transgene. Stable integration of the introduced DNA molecule into one of the host cell chromosomes is the simplest strategy for achieving this. However, genotoxicity resulting from random insertion of the transgene raises serious safety concerns that must be addressed if gene therapy is to enter the clinical mainstream. The following method makes use of the Rep integrase of adeno-associated virus to insert a transgene into the human AAVS1 site, a known "safe harbor" region within the human genome. This approach has the potential for application to novel gene therapy strategies for improved safety. In addition, with this method it is also possible to create cell lines carrying BAC transgenes in the AAVS1 site.

Original languageEnglish
Title of host publicationBacterial Artificial Chromosomes
EditorsKumaran Narayanan
Place of PublicationNY USA
PublisherSpringer
Chapter17
Pages309-321
Number of pages13
Volume1227
Edition2nd
ISBN (Electronic)9781493916528
ISBN (Print)9781493916511
DOIs
Publication statusPublished - 1 Jan 2015
Externally publishedYes

Publication series

NameMethods in Molecular Biology
PublisherHumana Press
ISSN (Print)1064-3745

Keywords

  • Adeno-associated virus
  • Gene therapy
  • Genotoxicity
  • Rep integrase
  • Site-specific integration

Cite this

McColl, B., Howden, S., & Vadolas, J. (2015). Site-specific integration of bacterial artificial chromosomes into human cells. In K. Narayanan (Ed.), Bacterial Artificial Chromosomes (2nd ed., Vol. 1227, pp. 309-321). (Methods in Molecular Biology). Springer. https://doi.org/10.1007/978-1-4939-1652-8_17