历史上,哺乳动物细胞的遗传分析需要分离具有改变特性的随机产生的突变细胞系,然后进行费力的遗传作图实验,以鉴定负责表型的突变基因。近年来,体细胞遗传学已经被功能基因组学筛选彻底改变了,每个蛋白质编码基因的表达都被系统地干扰,并确定扰动细胞的表型。我们在这里概述了一种新的功能基因组学筛选策略,该策略与常用方法根本不同。在这个战略中,我们表达了人工跨膜蛋白的文库,命名为捕获体,并选择具有所需表型的稀有细胞,因为,偶然,捕获体特异性干扰靶蛋白的表达或活性。然后从选定的细胞中回收活性捕获体,并可用作解剖研究中的生物过程的工具。我们还简要描述了我们如何使用这种新策略来提供对人类乳头瘤病毒进入细胞的复杂过程的见解。
Historically, the genetic analysis of mammalian cells entailed the isolation of randomly arising mutant cell lines with altered properties, followed by laborious genetic mapping experiments to identify the mutant gene responsible for the phenotype. In recent years, somatic cell genetics has been revolutionized by functional genomics screens, in which expression of every protein-coding gene is systematically perturbed, and the phenotype of the perturbed cells is determined. We outline here a novel functional genomics screening strategy that differs fundamentally from commonly used approaches. In this strategy, we express libraries of artificial transmembrane proteins named traptamers and select rare cells with the desired phenotype because, by chance, a traptamer specifically perturbs the expression or activity of a target protein. Active traptamers are then recovered from the selected cells and can be used as tools to dissect the biological process under
study. We also briefly describe how we have used this new strategy to provide insights into the complex process by which human papillomaviruses enter cells.