A genotype-phenotype screening system using conditionally immortalized immature dendritic cells

STAR Protoc. 2021 Aug 12;2(3):100732. doi: 10.1016/j.xpro.2021.100732. eCollection 2021 Sep 17.


Here, we describe a protocol for CRISPR/Cas9-mediated gene knockout in conditionally immortalized immature dendritic cells (DCs), which can be limitlessly expanded before differentiation. This facilitates the genetic screening of DC functions in vitro including assessment of phagocytosis, cytokine production, expression of co-stimulatory or co-inhibitory molecules, and antigen presentation, as well as evaluation of the capacity to elicit anticancer immune responses in vivo. Altogether, these approaches described in this protocol allow investigators to link the genotype of DCs to their phenotype. For complete details on the use and execution of this protocol, please refer to Le Naour et al. (2020).

Keywords: CRISPR; cell biology; cell culture; high-throughput screening; immunology; model organisms.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antigen Presentation / physiology
  • CRISPR-Cas Systems
  • Cell Culture Techniques / methods*
  • Cell Differentiation / physiology
  • Cell- and Tissue-Based Therapy
  • Cells, Cultured
  • Cytokines / metabolism
  • Dendritic Cells / classification*
  • Dendritic Cells / cytology*
  • Dendritic Cells / immunology
  • Dendritic Cells / physiology
  • Female
  • Gene Knockout Techniques / methods*
  • Genotype
  • Humans
  • Mice
  • Mice, Inbred C57BL
  • Phagocytosis / physiology
  • Phenotype


  • Cytokines