The CD300LD Knockout HCT 116 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal population derived from HCT 116 colorectal carcinoma cells, with targeted disruption of the CD300LD gene. This loss-of-function model provides a heterogeneous pool of edited cells, enabling functional studies while accounting for population variability. The polyclonal format is well-suited for screening applications and multiple assay types, avoiding biases introduced by single-cell cloning. Gene inactivation is achieved via CRISPR/Cas9, and the cells can be immediately analyzed by techniques such as Western blotting and flow cytometry.
The HCT 116 host cell line is a human colorectal carcinoma epithelial line characterized by microsatellite instability (MSI-H), a KRAS G13D mutation, and MLH1 deficiency. These cells are widely employed in cancer research due to their well-defined genetic background and robust growth characteristics. Their epithelial origin and tumorigenic properties make them a relevant model for colorectal cancer signaling, drug sensitivity testing, and studies of tumor-immune interactions. The HCT 116 line supports both in vitro and in vivo experimentation.
CD300LD is an activating immune receptor that signals through ITAM-bearing adaptors Fc??RI?? and DAP12. Upstream activation by IgE crosslinking, stem cell factor (SCF), IL-3, or PGE2 triggers recruitment and phosphorylation of spleen tyrosine kinase (Syk) and linker for activation of T cells (LAT). This initiates a pathway involving phospholipase C??1 (PLC??1)-dependent calcium flux and transcription factors NFAT and NF-??B, leading to mast cell and basophil degranulation and release of pro-inflammatory cytokines including IL-4, IL-13, and TNF-??. Although primarily characterized in allergic effector cells, CD300LD may also function in epithelial tumor contexts, making its knockout in HCT 116 cells a valuable tool for exploring non-classical roles.
In HCT 116 colorectal cancer cells, CD300LD knockout enables dissection of putative immune receptor signaling within a tumor epithelial setting. The interplay between CD300LD-mediated pathways and the oncogenic KRAS/MLH1-deficient background can be examined, potentially revealing contributions to proliferation, migration, or cytokine production. Co-culture assays with mast cells or basophils can assess how CD300LD deficiency in tumor cells modulates immune cell activation and cytokine release, offering insights into tumor-immune crosstalk relevant to colorectal cancer progression.
Key research applications include studying immune-related signaling in colorectal cancer, screening therapeutic antibodies targeting CD300LD, and functional analyses in allergic disease models. Standard validation assays such as Western blotting, RT-qPCR, and flow cytometry confirm gene disruption, while MTT/BrdU proliferation, migration/invasion, and cytokine ELISA assays enable phenotypic characterization. The polyclonal pool supports high-throughput screening and drug discovery targeting CD300LD. For further technical information, please contact Ascent Research.
