2nd Edition of Cell & Gene Therapy World Conference 2026

Abstract

Cellular senescence is driven by progressive epigenetic dysregulation and represents a pivotal target for anti-aging intervention. Conventional reprogramming strategies depend on exogenous transgene delivery, which entails risks of genomic insertion and aberrant expression. Here, we developed MultiTap-CRISPRa, a novel epigenetic reprogramming platform based on tandem gRNA arrays for synchronized, potent and multiplexed activation of endogenous genes. Using Golden Gate and Gibson assembly, we constructed optimized tandem gRNA vectors targeting OCT4, SOX2 and KLF4 (OSK) promoter regions. Following systematic gRNA screening and quantitative PCR validation, this system elicited robust multiplex gene activation: OCT4 11.82-fold, SOX2 5.96-fold, and KLF4 9.56-fold in human cells. We further established a Zeocin-induced cellular senescence model and confirmed that MultiTap-CRISPRa remains highly functional in senescent cells, enabling efficient and stable OSK activation under stress conditions. Preliminary results suggest transient transfection is insufficient to trigger epigenetic resetting. We are currently developing a stable, long-term expression system to achieve sustained OSK activation and genuine epigenetic remodeling for cellular rejuvenation. MultiTap-CRISPRa provides a highly modular, safe and efficient tool for multiplex gene regulation and epigenetic reprogramming. This platform establishes a novel strategy for cellular senescence intervention and holds significant translational potential for anti-aging and rejuvenation therapies.