Core Viewpoint - The article discusses the emerging understanding of synonymous mutations in human cells, challenging the traditional view of these mutations as neutral and highlighting their potential impact on cellular adaptability and disease [2][5][6]. Group 1: Research Background - A study by a team from the University of Michigan suggested that synonymous mutations in yeast may not be neutral and could affect cellular adaptability, reigniting interest in their biological effects [1]. - Previous research has linked a small number of synonymous mutations to human diseases, indicating their potential role as cancer drivers, but experimental confirmation remains limited [6]. Group 2: New Research Findings - A new study published by researchers from Peking University developed a high-throughput screening technology named PRESENT to investigate functional synonymous mutations in the human genome [4]. - The research utilized an advanced prime editing system (PEmax) to create a library targeting 3,644 human protein-coding genes, allowing for large-scale screening of synonymous mutations [7]. Group 3: Methodology and Tools - The study integrated single-cell screening methods with the PRESENT technology, termed DIRECTED-seq, to systematically evaluate the impact of identified synonymous mutations on gene expression [8]. - A specialized machine learning model called DS Finder was developed to analyze the effects of functional synonymous mutations on various biological processes, such as mRNA splicing and transcription [9][11]. Group 4: Key Findings - The research indicated that synonymous mutations exhibit different fitness effects compared to non-synonymous mutations, although their phenotypic distribution was similar to negative controls [9]. - The study identified that synonymous mutations could alter RNA folding and affect translation, with PLK1_S2 being a notable example, and combined screening data with predictive models to identify clinically relevant synonymous mutations [9].