BTK Knockout HCT 116 Polyclonal Cells provide a CRISPR/Cas9-edited polyclonal knockout cell population for the study of Bruton??s tyrosine kinase (BTK) in a human colorectal carcinoma background. This heterogeneous pool of gene-disrupted HCT 116 cells is generated without single-cell cloning, enabling robust loss-of-function experiments without clonal bias. The polyclonal format allows researchers to interrogate BTK-dependent phenotypes across a population of knockout alleles, facilitating functional genomics, pathway analysis, and drug response studies in a well-characterized epithelial cancer model.
HCT 116 is a near-diploid, mismatch repair-deficient colorectal carcinoma cell line harboring mutant MLH1 and displaying high microsatellite instability (MSI-H), while retaining wild-type KRAS and TP53. These genetic features recapitulate a subset of colorectal cancers and contribute to the line??s widespread use in DNA repair, cancer signaling, and therapeutic response research. The epithelial origin and maintained signaling networks make HCT 116 particularly suitable for examining the role of BTK, a kinase traditionally associated with hematopoietic malignancies, in a solid tumor context.
BTK is a cytoplasmic non-receptor tyrosine kinase predominantly known for its essential function in B-cell receptor (BCR) signaling, where it is activated downstream of SRC-family kinases LYN and SYK and the adaptor BLNK (SLP-65). Activated BTK phosphorylates PLC??2, triggering calcium mobilization and downstream activation of the PI3K-AKT, NF-??B, and MAPK/ERK cascades. BTK also participates in signal transduction from Toll-like receptors, chemokine receptors such as CXCR4, and certain cytokine receptors (e.g., IL-5, IL-6). Key downstream effectors include AKT, ERK1/2, NF-??B, STAT3, the anti-apoptotic protein BCL-xL, and the cell cycle regulator cyclin D2. Additionally, BTK interacts with kinases like TEC and PKC??, and engages the CARMA1-BCL10-MALT1 complex to modulate NF-??B activation in specific contexts.
In HCT 116 colorectal carcinoma cells, BTK promotes proliferation and migration independent of B-cell lineage, underscoring its relevance in epithelial tumor biology. The polyclonal knockout model allows dissection of BTK-mediated oncogenic signals in an MSI-high, KRAS wild-type background, providing insights into how BTK loss rewires the PI3K-AKT, MAPK, and NF-??B pathways. This system is valuable for investigating cross-talk between kinase networks and DNA repair deficiencies, and for elucidating mechanisms of resistance to targeted agents.
This BTK knockout cell population is ideally suited for a wide array of experimental approaches: Western blotting and RT-qPCR to confirm BTK disruption, phospho-signaling profiling for p-BTK, p-PLC??2, p-ERK, and p-AKT, cell proliferation (MTT, EdU) and migration/invasion assays, drug sensitivity testing with ibrutinib or acalabrutinib, apoptosis analysis by Annexin V/PI, colony formation assays, and transcriptomic analysis via RNA-seq. Researchers can employ these cells for functional dissection of BTK in colorectal cancer, preclinical evaluation of BTK inhibitors, and pathway analysis in mismatch repair-deficient cancers. For additional information or technical assistance, please contact Ascent Research.
