The CD274 Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human HT29 colorectal adenocarcinoma epithelial cell line. This product disrupts the CD274 gene, which encodes the immune checkpoint ligand PD-L1, providing a loss-of-function model for studying tumor immune evasion. The polyclonal nature ensures a diverse pool of gene-edited cells, suitable for heterogeneous population analyses.
The HT29 cell line was established from a colorectal adenocarcinoma of a 44-year-old Caucasian female and is a standard model in cancer research. HT29 cells form polarized epithelial monolayers with tight junctions, making them valuable for studying epithelial barrier function and colorectal tumorigenesis. Their robust in vitro growth and well-characterized molecular profile underpin their utility in target-gene knockout studies.
CD274 encodes PD-L1, an immune inhibitory ligand that binds PD-1 (PDCD1) on T cells, triggering SHP2-mediated dephosphorylation of ZAP70 and LCK and dampening PI3K/AKT and ERK signaling. Transcriptionally regulated by IFN-?? via STAT1/STAT3 and by NF-??B, MYC, and HIF1A, PD-L1 also interacts with CD80. This checkpoint axis suppresses T cell activation, proliferation, and cytokine production, promoting immune evasion. In addition to the canonical PD-1 pathway, PD-L1 signaling intersects with JAK/STAT, PI3K/AKT, and MAPK/ERK cascades.
In HT29 cells, PD-L1 expression contributes to immune evasion, and its knockout permits investigation of T cell reactivation in co-culture. This model allows assessment of how loss of PD-L1 affects T cell proliferation, cytokine secretion, and cytotoxicity, as well as its impact on epithelial barrier integrity and tumorigenic potential. The polyclonal knockout population mirrors tumor heterogeneity, enhancing the physiological relevance of experimental outcomes.
These polyclonal knockout cells are designed for immuno-oncology research, including immune checkpoint blockade, T cell exhaustion, and tumor microenvironment studies. Standard assays such as flow cytometry, co-culture T cell activation, ELISA, western blotting, and RT-qPCR can be used to validate PD-L1 disruption and functional effects. The product is well-suited for high-throughput drug screening of PD-1/PD-L1 inhibitors and functional genomics projects. For ordering and technical information, contact Ascent Research.
