RRC ID 12714
Author Huang S, Kawabe Y, Ito A, Kamihira M.
Title Adeno-associated virus Rep-mediated targeting of integrase-defective retroviral vector DNA circles into human chromosome 19.
Journal Biochem. Biophys. Res. Commun.
Abstract Retroviral vectors have been employed in clinical trials for gene therapy owing to their relative large packaging capacity, alterable cell tropism, and chromosomal integration for stable transgene expression. However, uncontrollable integrations of transgenes are likely to cause safety issues, such as insertional mutagenesis. A targeted transgene integration system for retroviral vectors, therefore, is a straightforward way to address the insertional mutagenesis issue. Adeno-associated virus (AAV) is the only known virus capable of targeted integration in human cells. In the presence of AAV Rep proteins, plasmids possessing the p5 integration efficiency element (p5IEE) can be integrated into the AAV integration site (AAVS1) in the human genome. In this report, we describe a system that can target the circular DNA derived from non-integrating retroviral vectors to the AAVS1 site by utilizing the Rep/p5IEE integration mechanism. Our results showed that after G418 selection 30% of collected clones had retroviral DNA targeted at the AAVS1 site.
Volume 417(1)
Pages 78-83
Published 2012-1-6
DOI 10.1016/j.bbrc.2011.11.059
PII S0006-291X(11)02073-0
PMID 22138242
MeSH Chromosomes, Human, Pair 19 / genetics* DNA, Circular / genetics* DNA-Binding Proteins / genetics Dependovirus / genetics* Gene Targeting / methods* Genetic Vectors / genetics* HeLa Cells Humans Integrases / genetics* Mutagenesis, Insertional Recombination, Genetic Viral Proteins / genetics
IF 2.559
Times Cited 8
WOS Category BIOPHYSICS BIOCHEMISTRY & MOLECULAR BIOLOGY
Resource
DNA material pBluescript SKII+AV1-235-4491 (RDB01505)