The role of SIRT1 in regulation of transcription and splicing in mouse embryonic stem cells

SIRT1 is a nuclear enzyme that removes acetyl-groups from target proteins by using NAD+ as a co-substrate. SIRT1 regulates many vital biological processes such as metabolism, stress response, development, and aging. We have previously shown that SIRT1 is critically involved in mouse embryonic stem cell (mESC) maintenance and differentiation in part through modulation of the acetylation status of key transcription factors and their mediated amino acid metabolism and cell signaling. Recent reports have shown that a number of alternative splicing factors are deacetylation substrates of SIRT1. In this study, we aim to understand the total RNA transcriptome, including splicing landscapes, of WT and SIRT1 KO mESCs. Overall design: Total RNA was extracted from WT and SIRT1 KO mESCs cultured in ESGRO medium in triplicates. All RNA-seq libraries were prepared with the TruSeq Stranded/Ribo kit (Illumina, San Diego, CA) and sequenced using the pair-end 76bp protocol at about 520 million reads per library using the NovaSeq platform (Illumina) per the manufacturer's protocol.