The BCR knockout HCT 116 polyclonal cells are a CRISPR/Cas9-edited polyclonal knockout cell population designed to disrupt the human BCR gene in the HCT 116 colorectal carcinoma cell line. This loss-of-function model leverages a heterogeneous pool of edited cells, enabling the study of BCR-dependent phenotypes without clonal selection. The polyclonal format preserves genetic diversity and mitigates concerns about clonal artifacts, making it suitable for pooled functional genomics and phenotypic screening approaches.
The HCT 116 cell line originates from a male patient with colorectal adenocarcinoma and was established from ascites fluid at a metastatic site. It exhibits a near-diploid karyotype and is widely employed as a model system for colorectal cancer biology, including investigations into tumor signaling, drug sensitivity, and metastatic mechanisms. Its genetic stability and well-characterized signaling landscape render it a robust platform for interrogating gene function in a relevant epithelial tumor context.
BCR encodes a multidomain protein that functions as both a serine/threonine kinase and a Rho GTPase-activating protein (GAP) for the small GTPases RAC1, CDC42, and RAP1. Through its GAP domain, BCR accelerates GTP hydrolysis, thereby negatively regulating these GTPases and attenuating their downstream signaling cascades. Under normal conditions, BCR suppresses RAC1- and CDC42-driven cytoskeletal reorganization, cell migration, and adhesion. BCR ablation leads to sustained activation of these GTPases, promoting downstream effectors such as the PAK kinases, NF-??B, and the MAPK pathways (ERK, JNK, p38). Additionally, BCR interacts with adaptor proteins GRB2, SOS1, and CRKL, and is regulated by upstream signals from integrins, cytokines, and SRC family kinases. In the context of hematopoietic malignancies, BCR is well known as the fusion partner of ABL1 in the oncogenic BCR-ABL tyrosine kinase, but its tumor-suppressive roles in solid tumors are under active investigation.
In colorectal adenocarcinoma, BCR’s function remains poorly defined, but its capacity to dampen Rho GTPase signaling suggests a potential tumor-suppressive role by restraining cell motility and invasive growth. The BCR knockout HCT 116 polyclonal cells offer a unique tool to explore how loss of BCR-mediated GTPase regulation contributes to enhanced migration, invasion, and proliferative signaling in a colorectal cancer background. This model enables researchers to dissect context-dependent functions of BCR, including its interplay with integrin and mTOR pathways, and to evaluate whether BCR loss sensitizes cells to chemotherapeutic agents or targeted therapies.
Researchers can employ these polyclonal knockout cells in a variety of assays, including western blotting for BCR and downstream targets, Rho GTPase activation pull-down assays to quantify RAC1 and CDC42 activity, and Boyden chamber migration and invasion assays to assess motility. Proliferation can be monitored via MTT or BrdU incorporation, while immunofluorescence microscopy reveals actin cytoskeletal changes. Transcriptomic profiling by RNA-seq and phospho-signaling analysis enable comprehensive characterization of pathway alterations. This product is ideal for investigating BCR-mediated GTPase regulation in colorectal cancer, dissecting mechanisms of metastasis, and testing responses to therapeutics. For further information about this knockout cell model or to place an order, please contact Ascent Research.
