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Endogenous Factors Causative of Spontaneous DNA Damage that Leads to Random Integration in Human Cells | OMICS International | Abstract
ISSN: 2329-6682

Gene Technology
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Research Article

Endogenous Factors Causative of Spontaneous DNA Damage that Leads to Random Integration in Human Cells

Haruna Kamekawa1, Aya Kurosawa1, Masumi Umehara1, Eriko Toyoda1 and Noritaka Adachi1,2*
1Graduate School of Nanobioscience, Yokohama City University, Yokohama 236-0027, Japan
2Advanced Medical Research Center, Yokohama City University, Yokohama 236-0004, Japan
Corresponding Author : Noritaka Adachi
Graduate School of Nanobioscience
Yokohama City University, Yokohama 236-0027, Japan
Tel: 81-45-787-2228
Fax: 81-45-787-2228
E-mail: [email protected]
Received August 29, 2013; Accepted October 01, 2013; Published October 03, 2013
Citation: Kamekawa H, Kurosawa A, Umehara M, Toyoda E, Adachi N (2013) Endogenous Factors Causative of Spontaneous DNA Damage that Leads to Random Integration in Human Cells. Gene Technology 2:105. doi:10.4172/2329-6682.1000105
Copyright: © 2013 Kamekawa H, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

Abstract

Random integration is a phenomenon in which transfected DNA molecules integrate into (random sites of) the host genome via non-homologous recombination. Although it is assumed that repair of DNA double-strand breaks leads to random integration events, how these endogenous DNA lesions are generated in living cells is poorly understood. In this study, we present evidence that DNA topoisomerase IIa (Top2α) and reactive oxygen species (ROS) are responsible for causing genomic DNA damage that leads to random integration. Specifically, we employed a human pre-B lymphocyte cell line to examine the effects of cellular Top2 expression levels and oxygen concentrations during cell culture. We find that treating cells with Top2α siRNA significantly reduces random integration frequency, while the absence of Top2β had little or no impact. We also show that cells continuously cultured under low (3%) oxygen culture conditions after electroporation display reduced random integration frequency compared to that under normal (21%) oxygen conditions. These findings support the notion that Top2α protein and ROS are endogenous factors that can produce DNA damage leading to random integration of transfected DNA in human cells.

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