The Biological Significance of Targeting Acetylation-mediated Gene Regulation for Designing New Mechanistic Tools and Potential Therapeutics Abstract: The dynamics between nucleosomal packaging and chromatin landscape regulate the transcriptional programming and biological outcomes of downstream genes. Epigenetic modifications play a central role in shaping the architecture of chromatin, which controls DNA accessibility by the transcriptional machinery. Acetylation of the lysine is one of the widespread epigenetic modifications that serves as a marker for gene activation, which intertwines the maintenance of cellular homeostasis and the regulation of signaling upon during stress. Besides, the biochemical horizon of acetylation ranges from orchestrating the stability and cellular localization of proteins that engage in the cell cycle, DNA repair, and metabolism. Further, lysine acetyltransferases (KATs) modulate the functions of transcription factors that govern cellular response to microbial infections, genotoxic stress, and inflammation. Due to its central role in many biological processes, mutations in KATs cause developmental and intellectual challenges and metabolic disorders. Despite the availability of tools for detecting acetylation, the mechanistic knowledge of the acetylation-mediated cellular processes remains limited. This review aims to integrate biochemical, molecular, and structural bases of KAT functions, which would help design tools for delineating acetylation-mediated mechanisms and increase our understanding of the biology of KATs towards developing disease treatment.