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Integration pattern of HIV-1 based lentiviral vector carrying recombinant coagulation factor VIII in Sk-Hep and 293T cells

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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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Authors and Affiliations

  1. Department of Clinical, Toxicological and Food Science Analysis, Faculty of Pharmaceutical Sciences of Ribeirão Preto, University of São Paulo, Ribeirão Preto, Brazil

    Elisa Maria de Sousa Russo-Carbolante

  2. Center for Cell Therapy and Regional Blood Center, Laboratory of Biotechnology, University of São Paulo, Ribeirão Preto, Brazil

    Elisa Maria de Sousa Russo-Carbolante, Virgínia Picanço-Castro, Daiani Cristina Cilão Alves, Andrielle Castilho Fernandes & Dimas Tadeu Covas

  3. Department of Animal Biotechnology, University of Nevada, Reno, Nevada, USA

    Graça Almeida-Porada

  4. DRK-Blutspendedienst Baden Württemberg—Hessen gGmbH, Institut für Transfusionsmedizin und Immunhämatologie, Klinikum der Johann Wolfgang Goethe Universität, Frankfurt am Main, Germany

    Torsten Tonn

  5. Department of Clinical Medicine, Faculty of Medicine of Ribeirão Preto, University of São Paulo, São Paulo, Brazil

    Dimas Tadeu Covas

  6. Departamento de Análises Clínicas Toxicológicas e Bromatológicas, Faculdade de Ciências Farmacêuticas de Ribeirão Preto, Av do Café s/n, Ribeirão Preto, SP, 14040-903, Brazil

    Elisa Maria de Sousa Russo-Carbolante

Authors
  1. Elisa Maria de Sousa Russo-Carbolante
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  2. Virgínia Picanço-Castro
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  3. Daiani Cristina Cilão Alves
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  4. Andrielle Castilho Fernandes
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  5. Graça Almeida-Porada
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  6. Torsten Tonn
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  7. Dimas Tadeu Covas
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Corresponding author

Correspondence to Elisa Maria de Sousa Russo-Carbolante.

Additional information

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

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