Effects of CRISPR-mediated knockout of Mitf, Tfe3 and Tfeb on the transcriptome of mouse BMDMs from WT and Grn KO animals [hex001130]

Previous single-cell RNA sequencing of microglia from aged WT and Grn KO mice revealed a microglial subset with transcriptomic signatures that were highly specific to Grn KO. Further epigenetic analysis demonstrated that these signatures were likely regulated by Mitf/Tfe family transcription factors. To investigate the necessity of these transcription factors in mediating the Grn KO transcriptomic response, we turned to an experimental in vitro validation. A multiplexed CRISPR approach was used to knockout Mitf, Tfe3 and Tfeb in BMDMs from WT or Grn KO mice. 10 days post-CRISPR, bulk RNA sequencing was performed to analyze the transcriptional profiles. These profiles identified the expected enrichment of Grn KO-specific genes in BMDMs from Grn KO mouse background. Strikingly, the levels of many of these genes were reversed with knockout of Mitf, Tfe3 and Tfeb.