The review discusses the emerging field of studying microproteins encoded by small open reading frames (sORFs) in the human genome. Despite being traditionally overlooked due to their small size, recent technological advancements such as ribosome profiling, mass spectrometry, and advanced computational approaches have led to the annotation of over 7000 sORFs. The review highlights the challenges in identifying functionally relevant microproteins and understanding their roles in different cellular contexts. It explores recent advancements in sORF research, focusing on new methodologies and computational approaches that have facilitated the identification and functional characterization of microproteins. The review also discusses the potential of these new tools in dissecting the diverse cellular roles of microproteins and their role in disease pathogenesis, paving the way for identifying new therapeutic targets. The introduction provides background on sORFs, their classification, and the challenges in their annotation. The review then delves into bioinformatic approaches, ribosome profiling, mass spectrometry-based proteomics, and genome editing using CRISPR for studying sORFs. Finally, it concludes by emphasizing the promise of these tools in advancing the field and the need for further investigations to elucidate the functional significance of microproteins.The review discusses the emerging field of studying microproteins encoded by small open reading frames (sORFs) in the human genome. Despite being traditionally overlooked due to their small size, recent technological advancements such as ribosome profiling, mass spectrometry, and advanced computational approaches have led to the annotation of over 7000 sORFs. The review highlights the challenges in identifying functionally relevant microproteins and understanding their roles in different cellular contexts. It explores recent advancements in sORF research, focusing on new methodologies and computational approaches that have facilitated the identification and functional characterization of microproteins. The review also discusses the potential of these new tools in dissecting the diverse cellular roles of microproteins and their role in disease pathogenesis, paving the way for identifying new therapeutic targets. The introduction provides background on sORFs, their classification, and the challenges in their annotation. The review then delves into bioinformatic approaches, ribosome profiling, mass spectrometry-based proteomics, and genome editing using CRISPR for studying sORFs. Finally, it concludes by emphasizing the promise of these tools in advancing the field and the need for further investigations to elucidate the functional significance of microproteins.