Department of Laboratory Medicine, Clinical Research Center, Karolinska Institutet, Karolinska University Hospital Huddinge, 141 86 Huddinge, Sweden.
Department of Molecular Biology and Functional Genomics, Stockholm University, 171 65 Stockholm, Sweden.
Molecules. 2019 Mar 12;24(5):1000. doi: 10.3390/molecules24051000.
, originally named c-, is an oncogene deregulated in many different forms of cancer. Translocation of the gene to an immunoglobulin gene leads to an overexpression and the development of Burkitt's lymphoma (BL). Sporadic BL constitutes one subgroup where one of the translocation sites is located at the 5'-vicinity of the two major promoters P₁ and P₂. A non-B-DNA forming sequence within this region has been reported with the ability to form an intramolecular triplex (H-DNA) or a G-quadruplex. We have examined triplex formation at this site first by using a 17 bp triplex-forming oligonucleotide (TFO) and a double strand DNA (dsDNA) target corresponding to the sequence. An antiparallel purine-motif triplex was detected using electrophoretic mobility shift assay. Furthermore, we probed for H-DNA formation using the BQQ-OP based triplex-specific cleavage assay, which indicated the formation of the structure in the supercoiled plasmid containing the corresponding region of the promoter. Targeting non-B-DNA structures has therapeutic potential; therefore, we investigated their influence on strand-invasion of anti-gene oligonucleotides (ON)s. We show that in vitro, non-B-DNA formation at the vicinity of the ON target site facilitates dsDNA strand-invasion of the anti-gene ONs.
，最初命名为 c-，是一种在许多不同形式的癌症中失调的癌基因。基因易位到免疫球蛋白基因导致过度表达和 Burkitt 淋巴瘤（BL）的发展。散发性 BL 构成了一个亚组，其中一个易位位点位于两个主要启动子 P₁和 P₂的 5'-附近。据报道，该区域内的一个非 B-DNA 形成序列具有形成分子内三链体（H-DNA）或 G-四链体的能力。我们首先使用 17 个碱基对的三链体形成寡核苷酸（TFO）和与 序列相对应的双链 DNA（dsDNA）靶标来检查该位点的三链体形成。通过电泳迁移率变动分析检测到反平行嘌呤基序三链体。此外，我们使用基于 BQQ-OP 的三链体特异性切割测定法探测 H-DNA 形成，这表明在含有相应 启动子区域的超螺旋质粒中形成了该结构。针对非 B-DNA 结构具有治疗潜力；因此，我们研究了它们对反基因寡核苷酸（ON）链入侵的影响。我们表明，在体外，ON 靶位点附近的非 B-DNA 形成促进了反基因 ON 的 dsDNA 链入侵。
