Precision Cell Fate Engineering with CRISPR Epigenetic Editing for Regeneration and Off-the-Shelf Immunotherapy
Abstract
Induced pluripotent stem cell (iPSC)-based regenerative medicine and cellular immunotherapy are severely hindered by heterogeneous cell phenotypes, unstable cellular functions and inconsistent large-scale manufacturing. Different from traditional CRISPR/Cas9 genome editing that induces DNA double-strand breaks and genomic risks, dCas9-mediated epigenetic editing offers a way to modulate gene regulation through targeted changes in DNA methylation and chromatin state without intentionally cutting the DNA. This review systematically summarizes core research progress and key findings in this field: targeted epigenetic modification mediated by CRISPRoff can establish persistent transcriptional memory during iPSC differentiation, supporting more stable control of cell state; IL1RN-targeted methylation editing of the promoter directly links epigenetic regulation to altered myeloid cell immune functions, providing evidence for a causal link between epigenetic state and cellular phenotype. The RENDER virus-like particle delivery platform may help address the cell-viability limitations associated with electroporation, realizing efficient and low-toxicity epigenetic editing in primary T cells. Combined with iPSC master cell bank technology, epigenetic editing may support more standardized expansion and batch-based production of CAR-T/CAR-NK off-the-shelf products. This review further clarifies existing limitations including off-target epigenetic effects, residual undifferentiated iPSCs and immune rejection, and proposes a systematic development strategy integrating precise epigenetic regulation, safe delivery and standardized iPSC manufacturing, providing a potential pathway for the clinical translation of cell fate engineering.
