Komarova E S, Dontsova O A, Pyshnyi D V, Kabilov M R, Sergiev P V
Institute of Functional Genomics, Lomonosov Moscow State University, Moscow, 119234 Russia.
Department of Chemistry, Lomonosov Moscow State University, Moscow, 119234 Russia.
Acta Naturae. 2022 Oct-Dec;14(4):20-37. doi: 10.32607/actanaturae.11820.
The Flow-seq method is based on using reporter construct libraries, where a certain element regulating the gene expression of fluorescent reporter proteins is represented in many thousands of variants. Reporter construct libraries are introduced into cells, sorted according to their fluorescence level, and then subjected to next-generation sequencing. Therefore, it turns out to be possible to identify patterns that determine the expression efficiency, based on tens and hundreds of thousands of reporter constructs in one experiment. This method has become common in evaluating the efficiency of protein synthesis simultaneously by multiple mRNA variants. However, its potential is not confined to this area. In the presented review, a comparative analysis of the Flow-seq method and other alternative approaches used for translation efficiency evaluation of mRNA was carried out; the features of its application and the results obtained by Flow-seq were also considered.
Flow-seq方法基于使用报告基因构建体文库，其中调控荧光报告蛋白基因表达的某个元件以成千上万种变体形式存在。将报告基因构建体文库导入细胞，根据其荧光水平进行分选，然后进行下一代测序。因此，在一次实验中基于成千上万的报告基因构建体就有可能识别出决定表达效率的模式。该方法在评估多个mRNA变体同时进行蛋白质合成的效率方面已变得很常见。然而，其潜力并不局限于此领域。在本综述中，对Flow-seq方法与用于评估mRNA翻译效率的其他替代方法进行了比较分析；还考虑了其应用特点以及通过Flow-seq获得的结果。
