CELL-SELEX on Hela cells - Raw sequence reads. Homo sapiens

Cell-based SELEX was performed using an RNA library containing a randomized 30-nt region flanked by fixed primer sequences. Cell-based selection was performed as previously described (Kim, Gu, 2013 PMID:23851587), by employing open PCR for DNA amplification during each selection round. Positive selection was performed on Hela cells transduced with a bicistronic lentiviral vector expressing the target surface receptor and GFP, while unmodified Hela cells, which lack expression of the target receptor, were used for negative selection. High throughput sequencing (HTS) was performed on the positive selection at rounds 0, 1, 3, 5, 6, 7, 8, and 9.

本研究采用带有固定引物侧翼序列的含随机化30核苷酸区域的RNA文库，开展基于细胞的指数富集配体系统进化（SELEX）技术实验。基于细胞的筛选流程参照已发表文献（Kim, Gu, 2013, PMID:23851587）开展，每一轮筛选阶段均采用开放式PCR进行DNA扩增。正向筛选以转染了表达目标表面受体与绿色荧光蛋白（GFP）的双顺反子慢病毒载体的海拉（Hela）细胞为靶标，而以未转染、不表达目标受体的野生型海拉细胞作为阴性筛选对照。分别对第0、1、3、5、6、7、8、9轮正向筛选产物进行高通量测序（HTS）。