One of the fundamental challenges in biology is understanding why genetically identical cells respond differently to the same stimulus. While some variation stems from genetic changes, even clonal cells can exhibit diverse behaviors. Our research addresses this by combining genomic and RNA biology to uncover both genetic and non-genetic sources of heterogeneity. Recent advances in Next Generation Sequencing (NGS) have revolutionised biomedicine, enabling rapid and affordable genome-wide analysis. However, while NGS readily identifies clonal mutations, resolving subclonal variation remains difficult. Genetic heterogeneity is a major driver of cancer initiation, progression, and relapse through drug-resistant clones. We are developing simple, cost-effective methods to detect mutations present in <1% of cells and to resolve complex structural variants, benchmarking these against clinical genomics standards. Beyond genetics, we investigate post-transcriptional RNA regulation and its role in cellular plasticity, transcriptional memory, and drug-tolerant persister states. Our lab pioneered 5PSeq, which exploits co-translational mRNA decay to provide genome-wide, drug-free ribosome dynamics profiles. Our lab uses a diversity of RNA genomics approaches (e.g., ribosome profiling, SLAM-Seq, isoform sequencing…) to dissect RNA biology in yeast, human samples, microbial communities, and antimicrobial resistance. Finally, our group also develops innovative epigenetic approaches to investigate chromatin heterogeneity at the single-cell level. The NAISS Medium allocation will support large-scale NGS method development and processing, and advanced modelling of translation–decay crosstalk. Deliverables include open pipelines, reference datasets linking decay signatures to cellular fitness, and diagnostic benchmarks for patient stratification and antimicrobial resistance detection.