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 (#201)

Hyuncheol Jung 1 2 3 , Pascal Devant 1 , Carter Ching 2 , Mineto Ota 1 4 5 , Emma Dann 1 4 5 , Ronghui Zhu 1 4 5 , Chandrima Modak 1 4 5 , Ana Vasquez-Ibarra 1 , Jennifer R Hamilton 6 7 , Zachary Steinhart 1 3 , Wayne Ngo 6 8 9 , Luis Sandoval 2 , Jae Hyung Jung 2 , Jae Hyun J. Lee 1 10 , Da Xu 7 , Meirui An 11 12 13 , Esha Urs 2 , Peixin Amy Chen 2 , Vincent Allain 1 2 14 , Takuya Tada 15 , Luke A Gilbert 16 17 18 , Brian R Shy 1 10 19 , Jonathan K Pritchard 4 5 , James K Nuñez 7 20 , Nathaniel R Landau 15 , David R Liu 11 13 21 , Justin Eyquem 1 2 3 18 19 22 23 , Jennifer A Doudna 1 6 7 8 9 24 25 26 , Jack Reid 2 , Alexander Marson 1 2 3 6 16 18 19 23 , Julia Carnevale 1 2 3 18 19
  1. Gladstone-UCSF Inst. for Genomic Immunology, San Francisco, CA
  2. Dept. of Medicine, Univ. of California San Francisco (UCSF), San Francisco, CA
  3. Parker Institute for Cancer Immunotherapy, San Francisco, CA
  4. Dept. of Genetics, Stanford Univ., Stanford, CA
  5. Dept. of Biology, Stanford Univ., Stanford, CA
  6. Innovative Genomics Institute, Univ. of California Berkeley, Berkeley, CA
  7. Dept. of Molecular and Cell Biology, Univ. of California Berkeley, Berkeley, CA
  8. Gladstone Inst. for Data Science and Biotechnology, San Francisco, CA
  9. California Inst. for Quantitative Biosciences, Univ. of California Berkeley, Berkeley, CA
  10. Dept. of Laboratory Medicine, UCSF, San Francisco, CA
  11. Merkin Inst., Broad Institute of MIT and Harvard, Cambridge, MA
  12. Dept. of Chemistry and Chemical Biology, Harvard Univ., Cambridge, MA
  13. Howard Hughes Medical Institute, Harvard Univ., Cambridge, MA
  14. Université Paris Cité, INSERM UMR1342, Hôpital Saint-Louis, Paris, France
  15. Dept. of Microbiology, NYU Grossman School of Medicine, New York, NY
  16. Arc Institute, Palo Alto, CA
  17. Dept. of Urology, UCSF, San Francisco, CA
  18. UCSF Helen Diller Family Comprehensive Cancer Center, San Francisco, CA
  19. Weill Cancer Hub West, Stanford Univ., Stanford, CA
  20. Chan Zuckerberg Biohub San Francisco, San Francisco, CA
  21. Dept. of Chemistry and Chemical Biology, Harvard Univ., Cambridge , MA
  22. Inst. for Human Genetics, UCSF, San Francisco, CA
  23. Dept. of Microbiology and Immunology, UCSF, San Francisco, CA
  24. Howard Hughes Medical Institute, Univ. of California Berkeley, Berkeley, CA
  25. Lawrence Berkeley National Laboratory, Berkeley, CA
  26. Dept. of Chemistry, Univ. of California Berkeley, Berkeley, CA

Primary human myeloid cells are promising candidates for immunotherapy, yet efficient and scalable technologies for genetic engineering and screening in these cells remain limited.  We found that the previously described CRISPR gene-editing method for human myeloid cells, which relies on electroporation, caused substantial cell loss and rendered the surviving cells functionally inert. Our lab has developed a virus-like particle (VLP)-based toolkit that delivers diverse CRISPR genome-editing modalities to human monocytes, macrophages, and dendritic cells with high efficiency, while preserving cell viability and innate immune responsiveness. VLP-mediated delivery of ribonucleoprotein payloads supports gene knockout, base editing, and epigenetic silencing. Furthermore, when combined with AAV-mediated donor delivery, this approach enables site-specific integration of large DNA sequences via homology-directed repair. We also developed SLICeVLP, a system combining sgRNA delivery via VPX-lentivirus with Cas9 protein delivery via engineered virus-like particles (eVLPs), and applied it to perform pooled loss-of-function screens and Perturb-seq in primary human macrophages. These methods have shown broad applicability to primary human dendritic cells as well, which will enable pooled loss- or gain-of-function screens in these cells in the future. Additionally, our efforts to engineer eVLPs with scFv-based binders to confer tropism to different myeloid cell subsets may allow us to utilize the eVLP system for in-vivo particle delivery, further expanding the discovery power and translational relevance of our tool set. Taken together, this platform enables unbiased functional genomics discovery in primary human myeloid cells, with direct implications for myeloid cell therapy design. We plan to use this technology to uncover genetic modulations that enhance dendritic cell function—such as cross-presentation and priming of the adaptive immune system—to improve anti-tumoral responses.