The IBTK Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal population with disruption of the endogenous IBTK locus in HEK293T cells. This polyclonal knockout model bypasses single-cell cloning, minimizing clonal artifacts and preserving population signaling dynamics. Loss of IBTK eliminates inhibition of Bruton tyrosine kinase (BTK) and disrupts Bcl-2 interaction, enabling unambiguous functional analysis in a well-characterized host background.
HEK293T is a widely used human embryonic kidney epithelial cell line, transformed with adenovirus 5 DNA and stably expressing SV40 large T antigen. This allows high-level episomal plasmid replication from SV40 origins, conferring exceptional transfectability and protein expression. Its robust growth and epithelial features make HEK293T an accessible platform for studying signal transduction, apoptosis, and transcriptional regulation across disciplines.
IBTK functions as a negative regulator of BTK kinase, a central component of B-cell receptor signaling. It directly binds BTK to suppress kinase activity and interacts with the anti-apoptotic protein Bcl-2 to modulate survival. IBTK stability is governed by the SCF ubiquitin ligase complex (including FBXO9) and the cAMP/PKA pathway. Downstream, IBTK restrains BTK-dependent phosphorylation of PLC??2, NF-??B activation, and MAP kinase cascades, thereby linking PI3K-Akt and apoptotic networks.
In HEK293T cells, which lack a complete BCR but express key downstream effectors like BTK and PLC??2, IBTK knockout enables dissection of BTK regulation and apoptosis crosstalk in a non-hematopoietic setting. IBTK disruption enhances BTK-mediated signaling and may sensitize cells to apoptotic stimuli, offering a controlled system to examine crosstalk between survival and death pathways. This model also facilitates study of IBTK??s role in transcriptional control and cell cycle regulation.
These polyclonal knockout cells support diverse molecular and functional assays. Western blotting and RT-qPCR confirm IBTK ablation, while co-immunoprecipitation verifies disrupted BTK or Bcl-2 interactions. Apoptosis assays (Annexin V, TUNEL) measure sensitization, and phospho-flow cytometry evaluates signaling flux. NF-??B reporter assays and cell viability tests with BTK inhibitors (e.g., ibrutinib) enable drug target validation. Applications extend to B-cell malignancy research, autoimmune disease studies, and fundamental signal transduction. For further details, please contact Ascent Research.
