Palmer Ella
Clinical Sciences Centre, Hammersmith Hospital, London, UK.
Methods Mol Biol. 2011;706:1-12. doi: 10.1007/978-1-61737-970-3_1.
Cell-based microarrays were first described by Ziauddin and Sabatini in 2001 as a novel method for performing high-throughput screens of gene function. They reported a technique whereby expression vectors containing the open reading frame (ORF) of human genes were printed onto glass microscope slides to form a microarray. Transfection reagents were added pre- or post-spotting and cells grown over the surface of the array. They demonstrated that cells growing in the immediate vicinity of the expression vectors underwent 'reverse transfection' and that subsequent alterations in cell function could then be detected by secondary assays performed on the array. Subsequent publications have adapted the technique to a variety of applications and have also shown that the approach works when arrays are fabricated using siRNAs and compounds. The potential of this method for performing analyses of gene function and identification of novel therapeutic agents has now been clearly demonstrated. Current efforts are focused on improving and harnessing this technology for high-throughput screening applications.
基于细胞的微阵列最早由齐亚丁和萨巴蒂尼于2001年描述为一种用于进行基因功能高通量筛选的新方法。他们报告了一种技术,即将含有人类基因开放阅读框(ORF)的表达载体打印在玻璃显微镜载玻片上以形成微阵列。在点样前或点样后添加转染试剂,然后让细胞在阵列表面生长。他们证明,在表达载体紧邻区域生长的细胞会发生“反向转染”,随后细胞功能的改变可以通过在阵列上进行的二次检测来检测。后续的出版物已将该技术应用于各种用途,并且还表明当使用小干扰RNA(siRNAs)和化合物制造阵列时该方法也有效。现在已经清楚地证明了这种方法在进行基因功能分析和鉴定新型治疗药物方面的潜力。目前的工作重点是改进并利用该技术用于高通量筛选应用。