FIGURE 2.

Overview of the commonly used techniques to identify and characterize sORFs and their encoded peptides. Novel sORFs and their products can be detected by the prediction algorithms using bioinformatic approaches, by generating peptide databases using improved mass spectrometry-based assays and by using ribosome profiling and related sequencing techniques to obtain translationally active transcripts. The predicted SEPs can be validated by various assays such as reporter-based overexpression, epitope tagging etc. Loss of function assays could be done to assess the cellular function of these SEPs.