基因治疗药物无义突变通读活性检测方法的研究进展

30%人类致病基因突变属无义突变，无义突变可导致众多疾病。无义突变通读剂具有无义突变通读活性，可诱导基因恢复其功能，属于基因治疗药物。基因编码区发生的无义突变可致蛋白翻译在无义突变位点提前终止，导致蛋白功能缺失，引起包括遗传病、肿瘤在内的众多疾病。少数化合物具有无义突变通读作用，可诱导全长蛋白翻译，是一种潜在的治疗药物。目前通读剂活性检测方法主要包括蛋白质截短试验和报告基因系统两大类型。本文回顾了无义突变通读活性检测方法的发展及最新进展，为药物高通量筛选方法的建立提供参考。

Nonsense mutation accounts for about 30% of human disease-causing alleles and is one of most frequent cause of many diseases. Nonsense mutation readthrough compounds,a kind of gene therapy drugs,can read through nonsense mutations and allow translation of full-length functional protein. Nonsense mutation in gene coding region can prematurely terminate translation and result in the loss of protein function,which is responsible for many diseases including genetic diseases and cancer. Some compounds have demonstrated readthrough ability and can induce translation of full-length protein,suggesting their possibility of being exploited as a potential therapy. To date,the most useful screening assays for readthrough compounds include protein truncation test and fluorescence reporter assay. Here,we review the protein truncation test and fluorescence reporter assay and describe the pros and cons of each method,providing information for the establishment of high throughput screening assay.