The AKAP13 Knockout A-549 Polyclonal Cells product provides a heterogeneous population of CRISPR/Cas9-edited A-549 cells carrying targeted disruption of the AKAP13 gene. This polyclonal knockout cell population is generated using CRISPR/Cas9-mediated gene editing to introduce loss-of-function alleles, creating a robust model for investigating AKAP13-dependent signaling without clonal selection. The resulting pool of edited cells enables the study of AKAP13 function in a genetically diverse background, making it suitable for experiments that require averaging across multiple knockout genotypes or for use in pooled functional screens.
The parental A-549 cell line is a widely used human lung adenocarcinoma epithelial model originally isolated from the tumor tissue of a 58-year-old male. These cells exhibit adherent epithelial morphology and retain key characteristics of alveolar basal epithelial cells, including the ability to form monolayers and respond to growth factors and cytokines relevant to lung cancer biology. A-549 cells are frequently employed in studies of non-small cell lung cancer (NSCLC) pathogenesis, drug resistance, and epithelial?Cmesenchymal transition (EMT).
AKAP13 (A-kinase anchoring protein 13) is a multifunctional scaffold protein that coordinates cAMP/PKA and Rho GTPase signaling pathways. It contains a PKA-anchoring domain that binds regulatory subunits of PKA, such as RI?? and RII??, and a DH-PH module that functions as a guanine nucleotide exchange factor (GEF) for RhoA. Upon activation by upstream signals including ??-adrenergic receptor agonists (e.g., isoproterenol), cAMP, and GPCR ligands, AKAP13 promotes RhoA-GTP loading, leading to downstream activation of effectors such as ROCK and SRF. This triggers transcriptional activation of genes like c-Fos, c-Jun, and YAP/TAZ, thereby influencing cytoskeletal dynamics, cell migration, and proliferation. AKAP13 also interacts with PKC, 14-3-3 proteins, and estrogen receptor ??, integrating multiple input signals.
Disruption of AKAP13 in A-549 cells provides a powerful tool to dissect the crosstalk between cAMP/PKA and Rho GTPase signaling in the context of lung adenocarcinoma. Given the role of AKAP13 in promoting cell migration and invasion, this knockout model can uncover how loss of AKAP13-dependent RhoA activation and downstream transcriptional programs affects tumor cell behavior. Moreover, as AKAP13 is implicated in cancer cell proliferation and survival, the polyclonal knockout cells enable the investigation of compensatory mechanisms and the identification of synthetic lethal interactions. This model is particularly relevant for studying the signaling networks that drive NSCLC metastasis and for evaluating therapeutic strategies targeting the cAMP/Rho axis.
Typical applications include western blotting and RT-qPCR to confirm AKAP13 disruption, RhoA activation assays to assess GTPase activity, wound healing and transwell invasion assays to measure migratory and invasive potential, and RNA-seq for transcriptomic profiling. The cells can be used to screen small-molecule inhibitors or biologics that modulate AKAP13-associated pathways, or to perform functional genomics studies such as CRISPRi/CRISPRa screens. For additional information, please contact Ascent Research.
