The AGRN Knockout A-549 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population derived from the A-549 human lung adenocarcinoma cell line. Designed to disrupt the AGRN gene, which encodes the heparan sulfate proteoglycan agrin, this model provides a heterogeneous gene-edited pool suitable for bulk functional studies without clonal selection bias. The polyclonal format enables robust assessment of agrin-dependent phenotypes in a therapeutically relevant lung cancer context, supporting pooled analyses and high-throughput screening applications.
The parental A-549 line, established from a 58-year-old Caucasian male with lung carcinoma, exhibits adherent epithelial morphology and harbors a KRAS G12S mutation, a frequent driver in non-small cell lung cancer. This cell line is a standard model for lung adenocarcinoma biology, drug response, and oncogenic signaling studies. The AGRN knockout in this KRAS-mutant background permits investigation of agrin??s contribution to tumor cell behavior and pathway interactions without confounds from divergent genetic backgrounds.
Agrin is a large proteoglycan critical for neuromuscular junction formation via the LRP4-MuSK-DOK7-RAPSN cascade, where it promotes acetylcholine receptor clustering. Beyond the synapse, agrin modulates cell adhesion and Hippo/YAP signaling: binding to LRP4 triggers MuSK autophosphorylation, leading to YAP and TAZ activation, cytoskeletal reorganization via PAK1 and LIMK, and integrin-mediated extracellular matrix engagement. Agrin also interacts with dystroglycan and ??-dystrobrevin, linking it to matrix adhesion. Thus, agrin integrates signals from LRP4, MuSK, integrins, and dystroglycan to regulate both synaptic assembly and cancer-associated transcription.
In A-549 cells, agrin potentiates YAP/TAZ-driven proliferation and migration, pathways frequently hijacked in lung adenocarcinoma. This polyclonal AGRN knockout model enables dissection of agrin-dependent YAP/TAZ activation, cell motility, and ECM remodeling in a KRAS-mutant background. Expression of LRP4, MuSK, and ??1 integrins in A-549 cells makes this model ideal for studying agrin-mediated crosstalk between Hippo/YAP and adhesion signaling. It also allows assessment of agrin??s impact on drug sensitivity, particularly to agents targeting YAP-regulated transcriptional programs.
This model supports diverse functional assays, including Western blot detection of AGRN, YAP, p-YAP, LRP4, and MuSK; immunofluorescence for YAP localization; cell viability (MTT/CCK-8) and Transwell migration/invasion assays; RT-qPCR and RNA-seq transcriptomics; co-immunoprecipitation of AGRN-LRP4; and phospho-kinase profiling. These approaches characterize agrin-dependent phenotypes and signaling networks. For technical inquiries or ordering information, please contact Ascent Research.
