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Related Experiment Video

Updated: Oct 23, 2025

Preparation of Tumor Antigen-loaded Mature Dendritic Cells for Immunotherapy
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A genotype-phenotype screening system using conditionally immortalized immature dendritic cells.

Liwei Zhao1,2, Peng Liu1,2, Wei Xie1,2,3

  • 1Equipe labellisée par la Ligue contre le cancer, Université de Paris, Sorbonne Université, INSERM UMR1138, Centre de Recherche des Cordeliers, Paris, France.

STAR Protocols
|August 25, 2021
PubMed
Summary

This study presents a new CRISPR/Cas9 gene knockout protocol for dendritic cells (DCs). This method enables comprehensive genetic screening of DC functions and their role in anticancer immunity.

Keywords:
CRISPRcell biologycell culturehigh-throughput screeningimmunologymodel organisms

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Area of Science:

  • Immunology
  • Molecular Biology
  • Cell Biology

Background:

  • Dendritic cells (DCs) are crucial for immune responses.
  • Genetic manipulation of DCs is essential for understanding their functions.
  • Existing methods for DC genetic studies have limitations.

Purpose of the Study:

  • To establish a protocol for CRISPR/Cas9-mediated gene knockout in immortalized immature dendritic cells (DCs).
  • To enable large-scale genetic screening of DC functions in vitro and in vivo.
  • To link DC genotype to phenotype for a deeper understanding of their roles.

Main Methods:

  • CRISPR/Cas9 gene editing technology.
  • Use of conditionally immortalized immature dendritic cells (DCs) for limitless expansion.
  • In vitro assays for phagocytosis, cytokine production, co-stimulatory/co-inhibitory molecule expression, and antigen presentation.
  • In vivo evaluation of anticancer immune responses.

Main Results:

  • A robust protocol for gene knockout in immortalized immature DCs was developed.
  • The protocol facilitates comprehensive functional screening of DCs.
  • The method allows for the direct correlation of DC genotype with observed phenotypes.

Conclusions:

  • This protocol significantly advances the genetic study of dendritic cells.
  • It provides a powerful tool for investigating DC biology and their role in immunity.
  • The described methods are valuable for researchers studying DC-mediated anticancer immune responses.