Castilho-Fernandes Andrielle, Fontes Aparecida Maria, Abraham Kuruvilla Joseph, de Freitas Marcela Cristina Corrêa, da Rosa Nathalia Gonsales, Picanço-Castro Virginia, de Sousa Russo-Carbolante Elisa Maria, Covas Dimas Tadeu
Faculty of Medicine of Ribeirão Preto, Center for Cell Therapy and Regional Blood Center, University of São Paulo, Ribeirão Preto, Brazil.
Biotechnol Lett. 2015 May;37(5):991-1001. doi: 10.1007/s10529-014-1764-2. Epub 2015 Feb 4.
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.
进行连接介导的聚合酶链反应（Ligation-mediated-PCR），随后分别对用携带重组人凝血因子IX（rhFIX）cDNA的嵌合载体转导的HepG2和Hek293细胞中的177个和150个整合位点进行定位。分析这些序列的染色体偏好性、CpG、转录起始位点（TSS）、重复元件、脆性位点和靶基因。在HepG2细胞中，rhFIX对6号和17号染色体的偏好性增加；到最近的CpG岛的中位距离为15,240个碱基对，37%的整合发生在RefSeq基因中。在Hek293细胞中，rhFIX对5号染色体的偏好性增加；到最近的CpG岛的中位距离为209,100个碱基对，74%的整合发生在RefSeq基因中。两种细胞系中的整合都远离转录起始位点。与该载体相关的整合模式在每个细胞系中都不同。
