cDNA expression cloning has been shown to be a powerful approach in the search for cellular factors that control virus replication. In this study, cDNA library screening using a pool of cDNA derived from interferon-treated human cells was combined with the MinION sequencer to identify cellular genes inhibiting Chikungunya virus (CHIKV) replication. Challenge infection of CHIKV to Vero cells transduced with the cDNA library produced virus-resistant cells. Then, the MinION sequence of cDNAs extracted from the surviving cells revealed that the open reading frames of TOM7, S100A16, N-terminally truncated form of ECI1 (ECI1ΔN59), and RPL29 were inserted in many of the cells. Importantly, the transient expression of TOM7, S100A16, and ECI1ΔN59 was found to inhibit the replication of CHIKV in Huh7 cells, indicating that these cellular factors were potentially anti-CHIKV molecules. Thus, our study demonstrated that cDNA expression cloning combined with the MinION sequencer allowed a rapid and comprehensive detection of cellular inhibitors against CHIKV.

在寻找控制病毒复制的细胞因子中，cDNA表达克隆已被证明是一种有效的方法。在这项研究中，使用来源于干扰素处理过的人类细胞的cDNA库与MinION测序仪结合进行cDNA文库筛选，以鉴定抑制基孔肯雅病毒（CHIKV）复制的细胞基因。挑战CHIKV感染用cDNA文库转导的Vero细胞产生的病毒抗性细胞。然后，从存活细胞中提取的cDNA的MinION序列显示，TOM7，S100A16，ECI1的N末端截短形式（ECI1ΔN59）和RPL29的开放阅读框已插入许多细胞中。重要的是，发现TOM7，S100A16和ECI1ΔN59的瞬时表达抑制了Huh7细胞中CHIKV的复制，表明这些细胞因子可能是抗CHIKV分子。因此，我们的研究表明，与MinION测序仪结合的cDNA表达克隆可快速，全面地检测针对CHIKV的细胞抑制剂。