Microproteins are defined as polypeptides of 100-150 or fewer amino acids. With the integrated application of ribosome profiling (Ribo-Seq), mass spectrometry, and bioinformatic approaches, more microproteins have been identified as being encoded by small open reading frames (sORFs). The majority of microproteins are evolutionarily young and may represent species-specific events. This review highlights the current methods and their challenges for identification and characterization of novel micr