The BCAR1 Knockout A-549 Polyclonal Cells product comprises a CRISPR/Cas9-edited polyclonal knockout cell population derived from the A-549 human lung adenocarcinoma cell line, engineered to disrupt the BCAR1 gene. This genetically modified model provides a powerful tool for investigating the scaffold functions of BCAR1 in integrin and growth factor signal transduction. The polyclonal knockout format ensures a heterogeneous population with disrupted target gene expression, suitable for bulk functional assays where clonal variability is averaged. Researchers can utilize these cells to dissect BCAR1-dependent mechanisms without the constraints of single-cell clonal effects, facilitating robust phenotypic screening in cancer biology contexts.
A-549 cells were originally established from the lung carcinoma tissue of a 58-year-old Caucasian male and exhibit adherent epithelial morphology. This line is widely employed as a model system for non-small cell lung cancer, particularly for studies focusing on metastasis, drug resistance, and signal transduction. A-549 cells possess active integrin-mediated adhesion and migration circuits, as well as responsive growth factor pathways, making them an ideal host for interrogating the biological roles of BCAR1. The parental line is well-characterized for its expression of key focal adhesion components and its sensitivity to FAK/SRC inhibitors, providing a relevant background for comparative knockout analyses.
BCAR1 (also known as p130Cas) is a mechanosensitive scaffold protein that integrates signals from integrins and receptor tyrosine kinases such as the EGF receptor. Upon cell adhesion or growth factor stimulation, BCAR1 is phosphorylated by FAK and SRC family kinases at multiple tyrosine residues within its substrate domain, creating docking sites for SH2 domain-containing proteins. Critically, BCAR1 recruits the adaptor Crk, which in turn activates the guanine nucleotide exchange factor C3G to promote GTP-loading of Rap1, leading to Rac-mediated actin polymerization and cell migration. Additionally, BCAR1 directly binds the p85 regulatory subunit of PI3K, enhancing AKT activation to support survival signaling and contribute to anti-estrogen resistance. This molecular network positions BCAR1 at a nexus controlling cell adhesion dynamics, directional motility, and anchorage-independent growth via downstream effectors including Rap1, Rac, PAK, MEK, ERK, and AKT.
In the A-549 lung adenocarcinoma context, BCAR1 knockout is anticipated to impair focal adhesion turnover and limit remodeling of the actin cytoskeleton, thereby attenuating migratory and invasive phenotypes. Because A-549 cells rely on integrin ??1 signaling for substrate attachment and motility, disruption of BCAR1??a core downstream transducer??enables dissection of integrin-to-actin signaling independently of upstream receptor perturbations. Furthermore, the model facilitates investigation of cross-talk between integrin and EGF receptor pathways, as BCAR1 integrates signals from both inputs. This is particularly relevant for understanding acquired resistance to targeted therapies, where enhanced BCAR1 signaling has been implicated in bypassing growth factor receptor inhibition.
This product is well-suited for a diverse array of experimental approaches. Typical functional assays include wound healing and transwell invasion to quantify migratory capacity, cell adhesion assays on extracellular matrix components, and immunofluorescence microscopy to visualize focal adhesion structures. Biochemical analyses encompass co-immunoprecipitation of BCAR1-Crk complexes, phospho-specific western blotting for BCAR1, and pharmacological challenges with FAK or SRC family kinase inhibitors to map signaling dependencies. Transcriptomic profiling by RNA-seq and phospho-proteomic workflows can further delineate pathway rewiring upon BCAR1 loss. These cells serve cancer biology, drug resistance, and functional genomics research programs. For detailed product information, please contact Ascent Research.
