The CD47 Knockout HCT 116 Polyclonal Cells product provides a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HCT 116 human colorectal carcinoma epithelial cell line. This loss-of-function model carries a targeted disruption of the CD47 gene, achieved through CRISPR/Cas9-mediated gene editing to ablate CD47 protein expression. These polyclonal knockout cells are a heterogeneous pool, reflecting a range of editing outcomes without clonal selection, enabling robust functional studies in a genetically defined background.
The parental HCT 116 cell line is a well-characterized human colorectal carcinoma model harboring an activating KRAS G13D mutation and wild-type TP53, combined with MLH1 deficiency that results in a microsatellite instability-high (MSI-H) phenotype. This genetic profile renders the cells mismatch repair-deficient and highly relevant for studying colorectal tumorigenesis and immune evasion mechanisms. HCT 116 cells are widely employed to investigate intestinal epithelial transformation and the tumor microenvironment, making them a suitable host for dissecting the role of immunomodulatory molecules such as CD47.
CD47 encodes a transmembrane protein that acts as a pivotal ‘don’t eat me’ signal by binding signal regulatory protein alpha (SIRP??) on macrophages and other myeloid cells. This interaction triggers phosphorylation of the immunoreceptor tyrosine-based inhibitory motifs of SIRP??, leading to recruitment and activation of SHP-1 and SHP-2 phosphatases, which in turn dephosphorylate phagocytic signaling components and inhibit engulfment. CD47 expression is transcriptionally driven by multiple upstream regulators including MYC, HIF-1??, and NF-??B, and is enhanced by inflammatory cytokines such as TNF-??. Beyond phagocytosis regulation, CD47 interacts with integrins ??v??3, ??IIb??3, and the matricellular protein thrombospondin-1 to modulate cell adhesion and migration, and can crosstalk with the PI3K/Akt signaling axis.
In the CD47 knockout HCT 116 polyclonal cell background, disruption of CD47 eliminates the dominant anti-phagocytic checkpoint, inherently enhancing susceptibility to macrophage-mediated phagocytosis. Given the MSI-H status and elevated mutational burden of HCT 116 cells, loss of CD47 may further expose the tumor cells to innate immune clearance and potentially reduce tumorigenic capacity. This engineered model thus recapitulates a critical vulnerability in colorectal cancer immune evasion and provides a platform to dissect CD47-dependent survival mechanisms in a clinically relevant genetic context.
These polyclonal knockout cells support diverse functional assays, including macrophage-tumor co-culture phagocytosis experiments visualized by live-cell imaging or quantified by flow cytometry. Researchers can confirm CD47 protein depletion via western blotting and immunofluorescence, and assess the impact on cellular behavior through proliferation and migration assays. The model is ideally suited for anti-CD47 antibody screening, validation of SIRP??-blocking reagents, and dissection of signaling events downstream of the CD47-SIRP?? axis. For further information, please contact Ascent Research.
