BTK Knockout 143B Polyclonal Cells are a genetically modified population derived from the 143B human osteosarcoma cell line, in which the BTK gene has been disrupted through CRISPR/Cas9-mediated editing. This polyclonal knockout pool provides a heterogeneous loss-of-function model of Bruton??s tyrosine kinase, a key mediator of B-cell receptor (BCR) signaling. The absence of clonal selection preserves a range of genomic edits, enabling studies of BTK-dependent processes without artifacts from single-cell cloning.
The 143B parental line is a TP53-mutant osteosarcoma model known for aggressive tumorigenic characteristics including rapid proliferation and metastatic potential. While BTK expression is typically restricted to hematopoietic cells, aberrant activation has been reported in osteosarcoma, where it may drive survival and invasive phenotypes. This cell line thus offers a relevant platform to investigate non-canonical BTK functions in bone cancer biology.
BTK operates as a cytoplasmic tyrosine kinase activated downstream of BCR engagement by SRC family kinases LYN and SYK. Upon membrane recruitment, BTK phosphorylates PLCG2, facilitated by scaffold protein BLNK, triggering calcium mobilization and subsequent activation of NF-??B and NFAT transcription factors via PKC. BTK also intersects with the PI3K?CAKT cascade through interactions with GAB2 and SH3BP5, forming a signalosome that governs transcriptional programs for proliferation and survival.
In osteosarcoma, BTK knockout permits dissection of the kinase??s contribution to tumor cell proliferation, migration, and drug response. Loss of BTK may attenuate PI3K?CAKT and NF-??B signaling, exposing vulnerabilities or compensatory pathways. The model is valuable for evaluating BTK inhibitor specificity, such as ibrutinib, and for validating antibodies targeting BTK in solid tumor contexts.
Applications include western blotting and RT-qPCR to confirm BTK ablation, phospho-protein analysis of downstream effectors (e.g., PLCG2, AKT), and cell viability assays for inhibitor screening. The knockout cells support studies on BCR signaling in a non-hematopoietic environment, functional analysis of BTK in bone cancer, and exploration of X-linked agammaglobulinemia-linked pathways. For inquiries, contact Ascent Research.
