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

STAR Protocols

卷:2

Sebastian Thieme¹  Shuai Zhang²  Peng Liu²  Guido Kroemer²  Oliver Kepp³  Wei Xie⁴  Liwei Zhao⁴  Laurence Zitvogel⁴ 

[1] Corresponding author;

[2] Metabolomics and Cell Biology Platforms, Gustave Roussy, Villejuif, France;

[3] Université Paris Sud, Paris Saclay, Faculty of Medicine, Kremlin Bicêtre, France;

[4] Equipe labellisée par la Ligue contre le cancer, Université de Paris, Sorbonne Université, INSERM UMR1138, Centre de Recherche des Cordeliers, Paris, France;

关键词: cell biology;    cell culture;    high-throughput screening;    immunology;    model organisms;    CRISPR;   

DOI  :  

来源: DOAJ

【 摘 要 】

Summary: 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).

【 授权许可】

Unknown