In Silico Design and Computational Evaluation of CRISPR-Cas9 sgRNAs Targeting Aging-Associated Genes TP53, FOXO3, and SIRT1

This paper examines the role of three aging-related genes -TP53,FOXO3 and SIRT1 in cellular senescence and lifespan. First, TP53 encodes the tumor suppressor protein p53, which regulates cellular activity such as cell cycle and prevents uncontrolled proliferation, ultimately affecting cellular aging and apoptosis (National Library of Medicine,2020). FOXO3 is a transcription factor that expands life span by regulating genes involved in stress resistance, DNA repair, and apoptosis (Weizmann Institute of Science, 2025). SIRT1 is an NAD+-dependent deacetylase that controls metabolic processes and involves DNA repair and aging regulation ( Science,2022). Using public genetic databases and the CHOPCHOP CRISPR design platform, this study computationally designed and evaluated optimal sgRNAs for future CRISPR-Cas9 knockout experiments targeting these genes. This study provides computationally optimized sgRNA candidates for future functional studies of aging-associated genes. Lastly, the results of this research can be used for future in vitro loss-of-function studies to better understand the molecular mechanisms of aging.