HLA-E Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal cell population derived from HT29 colorectal adenocarcinoma cells, with targeted disruption of the HLA-E gene. This gene-edited pool eliminates functional HLA-E expression while maintaining the heterogeneous characteristics of the parental line. The product provides a relevant model for investigating non-classical MHC class I-mediated immune regulation, particularly in NK cell biology and tumor immune evasion studies.
The HT29 cell line originates from a primary colorectal adenocarcinoma of a 44-year-old female and is a standard model for intestinal epithelial biology and colorectal cancer research. HT29 cells are capable of differentiation into enterocyte-like cells and are widely used to study drug absorption, metabolism, and epithelial barrier function. Their robust growth and genetic manipulation tolerance make them an ideal host for CRISPR-based gene editing to explore immune-related pathways.
HLA-E is a non-classical MHC class I molecule that presents leader peptides from classical MHC-I to CD94/NKG2 receptors on NK and T cells. Expression is induced by IFN-?? and TNF-??. HLA-E?C??2-microglobulin?Cpeptide complexes primarily bind inhibitory NKG2A/CD94, recruiting SHP-1 phosphatase to suppress Vav1-dependent signaling, blocking cytotoxicity and cytokine secretion. Activating signals via NKG2C/CD94 are context-dependent. TGF-?? and IL-10 modulate this system. This checkpoint maintains self-tolerance but is often hijacked by tumors to escape NK-mediated clearance.
In colorectal cancer, HT29 cells may exploit HLA-E to avoid NK cell attack. Knockout of HLA-E in this polyclonal population potentially removes a critical inhibitory signal, enhancing susceptibility to NK-mediated cytotoxicity. This model allows researchers to isolate HLA-E’s contribution to immune evasion and to investigate combinatorial strategies targeting both the HLA-E/NKG2A axis and other immune checkpoints. It also facilitates study of how colorectal cancer cells modulate NK cell responses in a physiologically relevant epithelial setting.
Key applications include co-culture with primary NK cells or NK cell lines to measure cytotoxicity via LDH release or flow-based killing assays, along with IFN-?? secretion ELISA. Surface HLA-E loss can be validated by flow cytometry, while RT-qPCR and Western blotting confirm gene disruption. These cells serve in drug screening for NKG2A-blocking antibodies or small molecules and in mechanistic studies of HLA-E regulatory networks under cytokine stimulation. For more information or to place an order, please contact Ascent Research.
