The BTK Knockout DLD-1 Polyclonal Cells represent a CRISPR/Cas9-mediated polyclonal knockout cell population generated from the DLD-1 human colorectal adenocarcinoma cell line. This product delivers a heterogeneous pool of cells carrying targeted disruptions in the BTK gene, providing a physiologically relevant loss-of-function model without clonal selection. The polyclonal format preserves genetic diversity while ablating BTK protein expression across the population, enabling robust functional studies in a colorectal cancer epithelial background.
DLD-1 is a widely employed colorectal cancer cell line isolated from a Duke??s type C colorectal adenocarcinoma. The cells harbor well-characterized oncogenic mutations in the tumor suppressor APC and the GTPase KRAS, which drive constitutive Wnt/??-catenin and MAPK pathway activation. These epithelial cells serve as a standard model for studying colorectal tumor biology, drug sensitivity, and signal transduction. Their adherent growth and reliable culture characteristics make them suitable for a broad range of downstream assays in cancer research.
BTK encodes Bruton??s tyrosine kinase, a non-receptor kinase central to B-cell receptor (BCR) signaling. Following BCR engagement, CD79A/B ITAMs are phosphorylated by LYN, recruiting SYK, which activates BTK. BTK, in complex with BLNK, phosphorylates PLCG2, triggering calcium flux and downstream NF-??B and MAPK/ERK pathways. In colorectal cancer, BTK may be activated non-canonically via Toll-like receptors or CXCR4, contributing to PI3K/AKT and NF-??B pro-survival signals. BTK directly phosphorylates PLCG2 and is required for AKT1 and ERK1/2 phosphorylation, as well as NFATC1 nuclear translocation. Key interactors include BLNK, GAB1, and PIK3R1, which fine-tune signal propagation.
Knockout of BTK in DLD-1 cells disrupts these signaling axes, leading to impaired phosphorylation of PLCG2 and AKT, attenuated NF-??B activation via the CARD11-BCL10-MALT1 complex, and reduced expression of NF-??B target genes. As a result, the polyclonal BTK knockout cells exhibit diminished cell survival and proliferation in vitro, offering a relevant context for studying BTK dependency in colorectal cancer with oncogenic APC and KRAS mutations. This model allows researchers to dissect the contribution of BTK to colorectal cancer cell autonomous signaling, independent of its well-known functions in B-cell malignancies, and to explore potential cross-talk between mutant KRAS-driven pathways and BTK-mediated signals.
The BTK Knockout DLD-1 Polyclonal Cells are suited for phospho-signaling arrays and Western blotting of downstream targets (phospho-PLCG2, phospho-AKT) to map BTK-dependent events. They enable drug target validation of BTK inhibitors in solid tumors using viability (MTT, CellTiter-Glo), colony formation, and apoptosis (Annexin V) assays. Additional applications include migration/invasion transwell assays, RT-qPCR profiling, and immunofluorescence localization. This model is valuable for functional genomics and deciphering non-canonical BTK functions beyond B cells. For more information, contact Ascent Research.
