Mechano-epigenetic enhancement of CRISPR activation of target genes for cell reprogramming [ATAC-Seq CS]

Here, we show that transient nuclear deformation not only drives global remodeling of chromatin accessibility, DNA methylation, and higher-order genome architecture but also reproducibly increasing accessibility at promoters of transcription factors, including Ascl1 and Oct4.

By coupling CRISPR-based transcriptional activation (CRISPRa) with guide RNAs designed to target these mechanically primed regions, we achieve enhanced activation of endogenous genes and improved fibroblast-to-neuron reprogramming efficiency. In addition, mechanically informed CRISPR activation of endogenous Oct4 induces a transient partial reprogramming state that further boosts reprogramming outcomes. Together, these findings establish a mechano-epigenetic strategy for overcoming chromatin barriers and provide a general framework for improving cell fate conversion.