Nature biotechnologyAug 18, 2026

Virus-like particles allow precise gene editing and combined CRISPR testing in primary human immune cells

CRISPR Gene Editing Weekly Brief ↗PubMed ↗DOI ↗

Updated Aug 19, 2026

Abstract

A novel virus-like particle (VLP)-based toolkit enables efficient CRISPR editing in primary human myeloid cells.

The toolkit delivers multiple CRISPR editing methods to human monocytes, macrophages, and dendritic cells while maintaining cell viability and immune responsiveness.
VLP-mediated delivery allows for gene knockout, base editing, and epigenetic silencing.
This approach also supports the integration of large DNA sequences through homology-directed repair using adeno-associated virus for donor delivery.
Pooled loss-of-function and Perturb-seq screens in human macrophages identified TNFAIP3 as a key regulator of inflammatory polarization.
Ablation of TNFAIP3 resulted in a proinflammatory state that resisted suppressive repolarization and increased cytotoxicity in chimeric antigen receptor macrophages.

Simplified

Primary human myeloid cells hold promise for immunotherapies, yet efficient, scalable technologies for engineering and screening in these cells remain limited. Here we present a virus-like particle (VLP)-based toolkit that delivers diverse CRISPR editing modalities to human monocytes, macrophages and dendritic cells with high efficiency while preserving viability and innate immune responsiveness. VLP-mediated delivery of ribonucleoproteins supports gene knockout, base editing and epigenetic silencing. Combined with adeno-associated virus-mediated donor delivery, this approach enables site-specific integration of large DNA sequences by homology-directed repair. We developed SLICeVLP, which pairs sgRNA delivery by VPX-lentivirus with Cas9 protein delivery by engineered VLPs, and used it for pooled loss-of-function and Perturb-seq screens in human macrophages. We uncovered regulators of tumor necrosis factor (TNF) and CD80 expression, converging on TNFAIP3 as a central regulator of inflammatory polarization. TNFAIP3 ablation drove a proinflammatory state resistant to suppressive repolarization and enhanced cytotoxicity in chimeric antigen receptor macrophages. This system enables unbiased functional genomics in primary human myeloid cells, with implications for myeloid cell therapy design.