Flash Talk & Poster Presentation 18th International Symposium on Dendritic Cells 2026

Virus-like particles enable targeted gene engineering and pooled CRISPR screening in primary human myeloid cells (#288)

Julia Carnevale 1 2 , Hyuncheol Jung 1 2 , Pascal Devant] 1 2 , Carter Ching 1 2 , Mineto Ota 2 , Emma Dann 2 , Ronghui Zhu 2 , Chandrima Modak 2 , Ana Vasquez-Ibarra 2 , Jennifer Hamilton 2 3 , Zachary Steinhart 1 2 , Wayne Ngo 2 3 , Luis Sandoval 1 2 , Jae Hyung Jung 1 2 , Jae Hyun J. Lee 2 , Da Xu 3 , Meirui An 4 , Esha Urs 1 2 , Peixin Amy Chen 1 2 , Vincent Allain 1 2 , Takuya Tada 5 , Luke A. Gilbert 1 , Brain R. Shy 1 2 , Jonathan Pritchard 6 , James K Nuñez 3 , Nathaniel R. Landau 5 , David R. Liu 4 , Justin Eyquem 1 2 , Jennifer Doudna 2 3 , Alexander Marson 1 2
  1. UCSF, San Francisco, CALIFORNIA, United States
  2. Gladstone Institutes, San Francisco, California, united states
  3. University of California, Berkeley, Berkeley, California, United States
  4. Broad Institute of MIT and Harvard, Cambridge, MA, United States
  5. NYU Grossman School of Medicine, New York, NY, USA
  6. Stanford, CA, USA

Dendritic cells and other primary human myeloid cells are compelling immunotherapy candidates, yet the tools to engineer and screen them at scale remain limited. We found that the established approach, electroporation of CRISPR ribonucleoproteins, leaves these cells functionally inert: electroporated macrophages and dendritic cells fail to mount normal innate inflammatory responses, and the accompanying cell loss makes pooled, large-scale forward genetic screens impractical. We therefore developed 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-delivered ribonucleoproteins support gene knockout, base editing, and epigenetic silencing, and combining them with AAV-mediated donor delivery enables site-specific integration of large transgenes by homology-directed repair. Building on this, we developed SLICeVLP, which pairs sgRNA delivery by Vpx-containing lentivirus with Cas9 protein delivery by engineered VLPs, making pooled loss-of-function and Perturb-seq screens tractable in primary human myeloid cells. Screening in primary human macrophages identified regulators of TNF and of the costimulatory molecule CD80, converging on TNFAIP3 as a central control point for inflammatory polarization; TNFAIP3 ablation produced a pro-inflammatory state resistant to suppressive repolarization, with enhanced cytotoxicity. Because the same delivery and editing modalities are efficient in dendritic cells, this platform extends unbiased functional genomics across myeloid lineages.