Biotechnology Letters

, Volume 33, Issue 1, pp 23–31

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

  • Elisa Maria de Sousa Russo-Carbolante
  • Virgínia Picanço-Castro
  • Daiani Cristina Cilão Alves
  • Andrielle Castilho Fernandes
  • Graça Almeida-Porada
  • Torsten Tonn
  • Dimas Tadeu Covas
Original Research Paper

DOI: 10.1007/s10529-010-0387-5

Cite this article as:
de Sousa Russo-Carbolante, E.M., Picanço-Castro, V., Alves, D.C.C. et al. Biotechnol Lett (2011) 33: 23. doi:10.1007/s10529-010-0387-5

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.

Keywords

Coagulation factor VIIILentiviral vectorIntegration sites

Supplementary material

10529_2010_387_MOESM1_ESM.docx (18 kb)
Supplementary material 1 (DOCX 18 kb)

Copyright information

© Springer Science+Business Media B.V. 2010

Authors and Affiliations

  • Elisa Maria de Sousa Russo-Carbolante
    • 1
    • 2
    • 6
  • Virgínia Picanço-Castro
    • 2
  • Daiani Cristina Cilão Alves
    • 2
  • Andrielle Castilho Fernandes
    • 2
  • Graça Almeida-Porada
    • 3
  • Torsten Tonn
    • 4
  • Dimas Tadeu Covas
    • 2
    • 5
  1. 1.Department of Clinical, Toxicological and Food Science Analysis, Faculty of Pharmaceutical Sciences of Ribeirão PretoUniversity of São PauloRibeirão PretoBrazil
  2. 2.Center for Cell Therapy and Regional Blood Center, Laboratory of BiotechnologyUniversity of São PauloRibeirão PretoBrazil
  3. 3.Department of Animal BiotechnologyUniversity of NevadaRenoUSA
  4. 4.DRK-Blutspendedienst Baden Württemberg—Hessen gGmbH, Institut für Transfusionsmedizin und ImmunhämatologieKlinikum der Johann Wolfgang Goethe UniversitätFrankfurt am MainGermany
  5. 5.Department of Clinical Medicine, Faculty of Medicine of Ribeirão PretoUniversity of São PauloSão PauloBrazil
  6. 6.Departamento de Análises Clínicas Toxicológicas e BromatológicasFaculdade de Ciências Farmacêuticas de Ribeirão PretoRibeirão PretoBrazil