Abstract
Ligation-mediated-PCR was performed followed by the mapping of 177 and 150 integration sites from HepG2 and Hek293 transduced with chimera vector carrying recombinant human Factor IX (rhFIX) cDNA, respectively. The sequences were analyzed for chromosome preference, CpG, transcription start site (TSS), repetitive elements, fragile sites and target genes. In HepG2, rhFIX was had an increased preference for chromosomes 6 and 17; the median distance to the nearest CpG islands was 15,240 base pairs and 37 % of the integrations occurred in RefSeq genes. In Hek293, rhFIX had an increased preference for chromosome 5; the median distance to the nearest CpG islands was 209,100 base pairs and 74 % of the integrations occurred in RefSeq genes. The integrations in both cell lines were distant from the TSS. The integration patterns associated with this vector are different in each cell line.
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Acknowledgments
Authors would like to thank: FINEP (Grant no 01.07.0652.00), FAPESP (Grant no 1998/14247-6), CNPq (Grant no 310619/2012-2 and 2008/57877-3). KJA also acknowledges financial support of the program Professor Visitante do Exterior of CAPES (Grant no 6327109) and Pesquisador Visitante Internacional of USP (Grant no 13.1.1318.17.3). Also, Sandra Navarro Bresciani for helping with the figures.
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Kuruvilla Joseph Abraham: Bolsista Professor Visitante/FMRP-USP.
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Castilho-Fernandes, A., Fontes, A.M., Abraham, K.J. et al. Significant differences in integration sites of Moloney murine leukemia virus/Moloney murine sarcoma virus retroviral vector carrying recombinant coagulation factor IX in two human cell lines. Biotechnol Lett 37, 991–1001 (2015). https://doi.org/10.1007/s10529-014-1764-2
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DOI: https://doi.org/10.1007/s10529-014-1764-2