The CD300LD Knockout KYSE-150 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population, providing targeted disruption of the CD300LD gene in the human KYSE-150 esophageal squamous cell carcinoma (ESCC) cell line. This loss-of-function model enables investigation of CD300LD, an activating immune receptor of the CD300 family, within a cancer-relevant context. The polyclonal format, generated by CRISPR/Cas9-mediated gene disruption, yields a heterogeneous edited population suitable for functional assays without clonal isolation. It is designed for studies of CD300LD-dependent signaling in cancer-immune interactions.
The KYSE-150 cell line was established from a poorly differentiated ESCC of the esophagus and displays epithelial morphology. Widely used in esophageal cancer research, it serves as a model for examining cancer cell biology, drug sensitivity, and oncogenic pathways. Engineering a CD300LD knockout in this background offers a physiologically relevant system to dissect gene function in an ESCC framework, where immune modulation and inflammatory signaling are pivotal.
CD300LD functions as an activating receptor that pairs with the adaptor protein TYROBP (DAP12). Upon engagement of ligands such as phosphatidylserine, pathogen-derived lipids, or apoptotic cells, CD300LD triggers TYROBP-dependent ITAM-mediated signaling, leading to activation of SYK kinase. Downstream, SYK orchestrates PI3K-AKT and MAPK/ERK pathways, culminating in NF-??B-mediated transcription of pro-inflammatory cytokines like TNF and IL-6, and promotion of phagocytosis. The receptor is also regulated by inflammatory cytokines including IFN-?? and TNF-??, embedding it in innate immune signaling networks.
Within esophageal squamous cell carcinoma, CD300LD may influence the tumor microenvironment by modulating myeloid cell phagocytosis and cytokine secretion. This knockout model enables dissection of DAP12-mediated signaling effects on tumor immunity, potentially revealing roles in immune evasion and the response to immunotherapy. The polyclonal population captures editing diversity, facilitating exploration of functional heterogeneity in CD300LD-dependent processes.
Applications include flow cytometry to verify CD300LD knockout, western blotting for TYROBP-SYK signaling analysis, and phagocytosis assays with fluorescent targets. Cytokine secretion can be quantified by ELISA, while co-cultures with immune cells assess tumor-immune crosstalk. Transcriptomic profiling via RNA-seq identifies global changes upon CD300LD loss. For technical inquiries, please contact Ascent Research.
