Integration site selection by HIV-based vectors in dividing and growth-arrested IMR-90 lung fibroblasts.
DNA integration is a defining step in the retroviral life cycle and the basis of stable gene transfer in retrovirus-based gene therapy. Previous studies of integration by HIV-based vectors have shown that integration is not random, but favored in active transcription units. Studies to date have focused on HIV integration in dividing cells, leaving open the question of whether integration target site selection might differ in nondividing cells. According to one idea, division of the host cell might be required for favored integration in transcription units, possibly as a result of chromatin remodeling during DNA replication. Here we have investigated this issue by comparing integration in dividing IMR-90 primary lung fibroblasts to integration in nondividing IMR-90 cells arrested in G1 by serum starvation and contact inhibition. We identified several differences in integration site selection in arrested versus dividing cells, including the frequency of integration in transcription units and in gene-rich regions. However, integration in nondividing cells was in fact more favored in transcription units, contrary to the idea that cell division was important for this bias. These data provide the first view of lentiviral integration in nondividing cells and help constrain models for the mechanism of favored integration in genes.
Ciuffi
Angela
A
Department of Microbiology, University of Pennsylvania School of Medicine, Philadelphia, 19104-6076, USA.
Mitchell
Richard S
RS
Hoffmann
Christian
C
Leipzig
Jeremy
J
Shinn
Paul
P
Ecker
Joseph R
JR
Bushman
Frederic D
FD
D002455:Cell Division / Q000235:genetics*; D002460:Cell Line; D003260:Contact Inhibition / Q000235:genetics*; D017077:Culture Media, Conditioned; D016895:Culture Media, Serum-Free / Q000009:adverse effects; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism / Q000821:virology*; D016193:G1 Phase / Q000235:genetics; D015967:Gene Expression Regulation, Viral / Q000502:physiology; D005822:Genetic Vectors; D006678:HIV / Q000235:genetics*; D006801:Humans; D008168:Lung / Q000166:cytology* / Q000378:metabolism / Q000821:virology*; D008969:Molecular Sequence Data; D014158:Transcription, Genetic / Q000502:physiology; D016662:Virus Integration / Q000235:genetics*
D017077:Culture Media, Conditioned; D016895:Culture Media, Serum-Free