Virus-like particles enable targeted gene engineering and pooled CRISPR screening in primary human myeloid cells

Nature Biotechnology
(2026) Cite this article

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.

This is a preview of subscription content, access via your institution

Access Nature and 54 other Nature Portfolio journals

Get Nature+, our best-value online-access subscription

Receive 12 print issues and online access

Prices may be subject to local taxes which are calculated during checkout

Raw sequencing data for RNA-seq, CRISPR screens and Perturb-seq were deposited to the Gene Expression Omnibus database under accession numbers GSE327424 and GSE327644. All other data are contained in the manuscript and its Supplementary Information. Source data are provided with this paper.

All custom code used to process data and generate data representations is available from a public GitHub repository (https://github.com/mangochiral/Macs_CRISPR_2026).

Blanchard, L., Mijacika, A. & Osorio, J. C. Targeting myeloid cells for cancer immunotherapy. Cancer Immunol. Res. 13, 1700–1715 (2025).

Article 
CAS 
PubMed 
PubMed Central 

Google Scholar 

Parvanian, S., Ge, X. & Garris, C. S. Recent developments in myeloid immune modulation in cancer therapy. Trends Cancer 11, 365–375 (2025).

Article 
CAS 
PubMed 

Google Scholar 

Cao, L. L. & Kagan, J. C. Targeting innate immune pathways for cancer immunotherapy. Immunity 56, 2206–2217 (2023).

Article 
CAS 
PubMed 
PubMed Central 

Google Scholar 

Source: Read the original article on www.nature.com