BCR Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population produced by targeted disruption of the BCR gene in the HT29 human colorectal adenocarcinoma cell line. This loss-of-function model is generated via CRISPR/Cas9-mediated gene editing, resulting in a heterogeneous pool of cells with BCR disruption. The polyclonal format minimizes clonal selection biases and preserves population-level genetic diversity, making it suitable for robust functional studies of BCR-dependent pathways without artifacts associated with single-cell-derived clones.
HT29 cells, isolated from the colorectal adenocarcinoma of a 44-year-old Caucasian female, grow as adherent epithelial cells and serve as a well-characterized model of intestinal epithelium. This cell line retains key features of colorectal cancer and is widely employed to investigate oncogenic signaling, drug responses, and epithelial biology. Its reproducible growth characteristics and defined genetic background provide a reliable platform for generating knockout derivatives.
The BCR gene encodes a multifunctional protein that acts as a GTPase-activating protein (GAP) for the Rho-family GTPases RAC1 and CDC42, accelerating GTP hydrolysis to terminate their signaling. BCR also possesses serine/threonine kinase activity and transduces signals downstream of ABL kinase and receptor tyrosine kinases. It interacts with adaptor proteins GRB2, CRKL, and 14-3-3, which modulate its subcellular localization and function. Loss of BCR disrupts its GAP activity, leading to sustained activation of RAC1 and CDC42. This hyperactivation propagates through downstream effectors PAK and LIMK, ultimately dysregulating actin cytoskeleton dynamics and impacting pathways such as Neurotrophin and PDGF signaling.
In HT29 colorectal adenocarcinoma cells, BCR knockout provides a physiologically relevant context to examine its GAP function independently of the BCR-ABL oncoprotein. This model allows dissection of BCR’s role in Rho GTPase regulation and actin remodeling within an epithelial cancer background, where processes like cell migration, invasion, and polarity are frequently altered. The knockout highlights BCR??s contribution to maintaining epithelial architecture and may reveal tumor-suppressive or oncogenic activities in colorectal carcinogenesis.
These polyclonal knockout cells are suitable for diverse experimental applications. Researchers can directly measure RAC1 and CDC42 activation using Rho GTPase activity assays, visualize F-actin organization by phalloidin staining, and quantify cell migration via Transwell and wound healing assays. Molecular changes are assessable through RT-qPCR and Western blotting. The product facilitates investigation of BCR-ABL-independent signaling and colorectal cancer cell signaling. For further details or technical support, please contact Ascent Research.
