Abstract
293T and Sk-Hep-1 cells were transduced with a replication-defective self-inactivating HIV-1 derived vector carrying FVIII cDNA. The genomic DNA was sequenced to reveal LTR/human genome junctions and integration sites. One hundred and thirty-two sequences matched human sequences, with an identity of at least 98%. The integration sites in 293T-FVIIIDB and in Sk-Hep-FVIIIDB cells were preferentially located in gene regions. The integrations in both cell lines were distant from the CpG islands and from the transcription start sites. A comparison between the two cell lines showed that the lentiviral-transduced DNA had the same preferred regions in the two different cell lines.
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Acknowledgments
We thank Adriana Aparecida Marques who helped with the sequencing. This work was supported by CNPq (Conselho Nacional de Desenvolvimento Tecnológico) and FAPESP (Fundação de Amparo a Pesquisa do Estado de São Paulo).
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Purpose of work We have examined lentiviral integration site selection in hepatocyte and kidney cell lines to see whether there are determinants of integration tendency other than the structure and biological characteristics of lentiviruses.
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de Sousa Russo-Carbolante, E.M., Picanço-Castro, V., Alves, D.C.C. et al. Integration pattern of HIV-1 based lentiviral vector carrying recombinant coagulation factor VIII in Sk-Hep and 293T cells. Biotechnol Lett 33, 23–31 (2011). https://doi.org/10.1007/s10529-010-0387-5
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DOI: https://doi.org/10.1007/s10529-010-0387-5