The KLKB1 Knockout HCT 116 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human colorectal carcinoma HCT 116 parental line. This product provides a heterogeneous pool of cells carrying targeted disruptions in the KLKB1 gene, designed to ablate plasma prekallikrein expression and function. The polyclonal format preserves genetic diversity while enabling robust loss-of-function studies, and it serves as a versatile tool for investigating the biological roles of plasma kallikrein in cancer and hemostasis-related pathways without the constraints of single-cell clonal selection.
The parental HCT 116 cell line is an epithelial cell model established from a male patient diagnosed with colorectal adenocarcinoma. These cells are characterized by a KRAS G13D driver mutation, MLH1 deficiency, and microsatellite instability-high (MSI-H) status, which collectively render them representative of a molecularly defined subset of colorectal tumors. HCT 116 cells are widely employed in cancer research due to their rapid proliferation and well-characterized signaling networks, making them a suitable host for modeling gene functions in the context of colorectal carcinogenesis and therapeutic screening.
KLKB1 encodes plasma prekallikrein, a serine protease that occupies a central position within the contact activation system. Following activation by factor XIIa, plasma kallikrein cleaves high-molecular-weight kininogen to release bradykinin, promoting inflammation and vasodilation. Active kallikrein also activates factor XII and converts plasminogen to plasmin, linking intrinsic coagulation, fibrinolysis, and complement cascades. The pathway is regulated by C1 esterase inhibitor and prolylcarboxypeptidase, and signaling through bradykinin B2 receptors connects kallikrein activity to renin-angiotensin and kinin-kallikrein interplay.
Disruption of KLKB1 in the HCT 116 background offers a platform to dissect the tumor-cell-intrinsic roles of plasma kallikrein in colorectal cancer biology. The contact activation system components, including high-molecular-weight kininogen and bradykinin receptors, are increasingly recognized for their contributions to tumor microenvironment remodeling, angiogenesis, and inflammatory signaling. This knockout model enables researchers to investigate whether loss of prekallikrein alters cancer cell proliferation, migration, or invasion, and to assess its impact on coagulation-dependent metastasis or bradykinin-mediated paracrine communication within MSI-H tumor ecosystems.
Typical experimental applications include functional assessment of KLKB1 through proliferation and transwell migration/invasion assays, complemented by western blotting and immunofluorescence for protein validation. Bradykinin release can be quantified via ELISA, while kallikrein enzymatic activity is measurable using chromogenic substrates. Coagulation profiles may be evaluated by activated partial thromboplastin time (aPTT) assays, and global transcriptomic changes can be characterized by RNA sequencing. These polyclonal knockout cells are valuable for screening small-molecule kallikrein inhibitors or studying crosstalk between hemostatic and oncogenic pathways. For further information, please contact Ascent Research.
