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Mol Ther. 2016 Mar;24(3):592-606. doi: 10.1038/mt.2016.11. Epub 2016 Jan 12.

Genome-wide Profiling Reveals Remarkable Parallels Between Insertion Site Selection Properties of the MLV Retrovirus and the piggyBac Transposon in Primary Human CD4(+) T Cells.

Author information

1
Institute of Computer Science, Martin Luther University Halle-Wittenberg, Wittenberg, Germany.
2
German Centre for Integrative Biodiversity Research (iDiv) Halle-Jena-Leipzig, Leipzig, Germany.
3
Max Delbrück Center for Molecular Medicine, Berlin, Germany.
4
Institute of Virology, Hannover Medical School, Hannover, Germany.
5
Division of Medical Biotechnology, Paul Ehrlich Institute, Langen, Germany.

Abstract

The inherent risks associated with vector insertion in gene therapy need to be carefully assessed. We analyzed the genome-wide distributions of Sleeping Beauty (SB) and piggyBac (PB) transposon insertions as well as MLV retrovirus and HIV lentivirus insertions in human CD4(+) T cells with respect to a panel of 40 chromatin states. The distribution of SB transposon insertions displayed the least deviation from random, while the PB transposon and the MLV retrovirus showed unexpected parallels across all chromatin states. Both MLV and PB insertions are enriched at transcriptional start sites (TSSs) and co-localize with BRD4-associated sites. We demonstrate physical interaction between the PB transposase and bromodomain and extraterminal domain proteins (including BRD4), suggesting convergent evolution of a tethering mechanism that directs integrating genetic elements into TSSs. We detect unequal biases across the four systems with respect to targeting genes whose deregulation has been previously linked to serious adverse events in gene therapy clinical trials. The SB transposon has the highest theoretical chance of targeting a safe harbor locus in the human genome. The data underscore the significance of vector choice to reduce the mutagenic load on cells in clinical applications.

PMID:
26755332
PMCID:
PMC4786924
DOI:
10.1038/mt.2016.11
[Indexed for MEDLINE]
Free PMC Article

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