The CD47 Knockout HT29 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population in which the endogenous CD47 gene has been disrupted in the human HT29 colon adenocarcinoma cell line. This polyclonal pool provides a heterogeneous loss-of-function model for investigating CD47-dependent signaling and immune regulatory mechanisms, without clonal selection. The knockout product format allows researchers to assess population-level responses to CD47 ablation, minimizing clone-specific artifacts. This gene-edited cell model is suitable for functional studies where complete knockout of CD47 is required to dissect its role in phagocytosis regulation and tumor immune evasion.
The parental HT29 cell line is a well-characterized epithelial colorectal adenocarcinoma model derived from a 44-year-old female patient. HT29 cells display an adherent, epithelial-like morphology and are capable of forming polarized monolayers with functional tight junctions, making them valuable for intestinal epithelial barrier studies. Additionally, these cells produce mucin and are widely utilized in drug absorption and metabolism research. In colorectal cancer research, HT29 serves as a robust platform for investigating tumor cell biology, signaling pathways, and therapeutic responses, particularly in the context of immune checkpoint modulation.
CD47 encodes a cell surface glycoprotein that functions as a ??don??t eat me?? signal by binding to signal regulatory protein alpha (SIRP??) on macrophages, inhibiting phagocytosis. This triggers SIRP?? phosphorylation and recruitment of SHP-1 and SHP-2 phosphatases, leading to inhibition of integrin activation, nitric oxide production, and cytoskeletal remodeling. Upstream regulators include HIF1A, TNF??, IL-6, and NF-??B. CD47 also interacts with integrins (??v??3, ??2??1), thrombospondin-1, and VEGFR2, modulating cell adhesion, migration, and angiogenesis. Overexpression of CD47 on cancer cells enables immune evasion by suppressing macrophage-mediated clearance, making it a critical immune checkpoint in the tumor microenvironment.
In the HT29 colorectal adenocarcinoma model, CD47 knockout provides a powerful system to dissect the role of CD47 in immune evasion and phagocytosis regulation within a human intestinal tumor context. Since HT29 cells endogenously express CD47 and are derived from a colorectal tumor, disrupting CD47 allows direct assessment of how loss of this ??don??t eat me?? signal affects interactions with macrophages and other immune cells. This model is particularly relevant for studying the CD47-SIRP?? axis in colorectal cancer, where CD47 overexpression is associated with poor prognosis. The polyclonal knockout population enables evaluation of heterogeneous responses that more closely mimic in vivo tumor diversity.
The CD47 Knockout HT29 Polyclonal Cells are ideally suited for research applications including cancer immunotherapy, immune checkpoint blockade, and anti-CD47 antibody therapy development. They can be employed in phagocytosis assays with primary macrophages, flow cytometry to confirm CD47 loss, western blotting, and immunofluorescence for localization. Cell viability and migration/invasion assays facilitate functional analyses, and these cells support in vivo xenograft tumor models to evaluate anti-CD47 treatment and tumor microenvironment modulation. For further details, please contact Ascent Research.
