Your browser doesn't support javascript.
loading
A genome-wide analysis of lentivector integration sites using targeted sequence capture and next generation sequencing technology.
Ustek, Duran; Sirma, Sema; Gumus, Ergun; Arikan, Muzaffer; Cakiris, Aris; Abaci, Neslihan; Mathew, Jaicy; Emrence, Zeliha; Azakli, Hulya; Cosan, Fulya; Cakar, Atilla; Parlak, Mahmut; Kursun, Olcay.
Afiliación
  • Ustek D; Department of Genetics, Istanbul University, Institute for Experimental Medicine, Istanbul 34093, Turkey. dustek@istanbul.edu.tr
Infect Genet Evol ; 12(7): 1349-54, 2012 Oct.
Article en En | MEDLINE | ID: mdl-22613802
ABSTRACT
One application of next-generation sequencing (NGS) is the targeted resequencing of interested genes which has not been used in viral integration site analysis of gene therapy applications. Here, we combined targeted sequence capture array and next generation sequencing to address the whole genome profiling of viral integration sites. Human 293T and K562 cells were transduced with a HIV-1 derived vector. A custom made DNA probe sets targeted pLVTHM vector used to capture lentiviral vector/human genome junctions. The captured DNA was sequenced using GS FLX platform. Seven thousand four hundred and eighty four human genome sequences flanking the long terminal repeats (LTR) of pLVTHM fragment sequences matched with an identity of at least 98% and minimum 50 bp criteria in both cells. In total, 203 unique integration sites were identified. The integrations in both cell lines were totally distant from the CpG islands and from the transcription start sites and preferentially located in introns. A comparison between the two cell lines showed that the lentiviral-transduced DNA does not have the same preferred regions in the two different cell lines.
Asunto(s)

Texto completo: 1 Colección: 01-internacional Banco de datos: MEDLINE Asunto principal: Genoma Humano / Integración Viral / Lentivirus Tipo de estudio: Prognostic_studies Límite: Humans Idioma: En Revista: Infect Genet Evol Asunto de la revista: BIOLOGIA / DOENCAS TRANSMISSIVEIS / GENETICA Año: 2012 Tipo del documento: Article País de afiliación: Turquía

Texto completo: 1 Colección: 01-internacional Banco de datos: MEDLINE Asunto principal: Genoma Humano / Integración Viral / Lentivirus Tipo de estudio: Prognostic_studies Límite: Humans Idioma: En Revista: Infect Genet Evol Asunto de la revista: BIOLOGIA / DOENCAS TRANSMISSIVEIS / GENETICA Año: 2012 Tipo del documento: Article País de afiliación: Turquía