The TSPAN1 Knockout A549 Cell Line is a CRISPR/Cas9-edited knockout cell line derived from A549 human lung adenocarcinoma cells. It provides a stable loss-of-function model for TSPAN1, generated by CRISPR/Cas9-mediated disruption of the TSPAN1 gene. This knockout cell line enables precise investigation of TSPAN1 roles in cell adhesion, migration, and invasion without transient silencing artifacts. Researchers can use this isogenic model to dissect TSPAN1-dependent signaling networks and perform rescue experiments. The product maintains the KRAS G12S mutation and wild-type TP53 of the parental A549 line.
The A549 cell line is an established model of human lung adenocarcinoma derived from a 58-year-old male, displaying properties of alveolar type II epithelial cells. It carries a KRAS G12S activating mutation and retains wild-type TP53, reflecting a common NSCLC genotype. A549 cells are widely used to study lung cancer cell adhesion, migration, and invasion, processes heavily dependent on integrin signaling. Their metastatic behavior and responsiveness to growth factors make them suitable for exploring tetraspanin-mediated regulation of tumor progression.
TSPAN1 is a tetraspanin that organizes integrin complexes within tetraspanin-enriched microdomains. It interacts with integrin ??3??1 and ??6??1, CD151, and CD9 to modulate adhesion, migration, and signal transduction. TSPAN1 is activated by TGF-?? and EGF, leading to downstream phosphorylation of AKT and ERK1/2, and upregulation of MMP-2 and MMP-9. Knockout disrupts these complexes, attenuates AKT/ERK signaling, reduces MMP activity, and alters EMT markers like E-cadherin and vimentin. The FAK-Src pathway, linked to integrin signaling, is also affected. The transcription factor Snail, a regulator of EMT, may be influenced by TSPAN1 loss.
In A549 cells, TSPAN1 drives metastatic behavior by sustaining integrin-based adhesion and ECM degradation. CRISPR/Cas9-mediated knockout in this KRAS-mutant background uncouples oncogenic KRAS from tetraspanin-organized integrin networks. This model allows assessment of TSPAN1 loss on invasiveness, EMT, and AKT/ERK signaling without animal genetic complexity. It is ideal for comparative studies between wild-type A549 and TSPAN1-null cells to define tetraspanin contributions to lung cancer metastasis. The cell line also facilitates drug screening for inhibitors targeting TSPAN1-enhanced signaling.
The TSPAN1 Knockout A549 Cell Line supports diverse research applications, including dissection of integrin signaling, EMT, and lung cancer metastasis. Typical assays include Transwell migration/invasion, wound healing, and cell adhesion to measure phenotypic changes. Western blotting and phospho-flow cytometry enable analysis of AKT and ERK activation, while RT-qPCR and RNA-seq profile EMT-related genes. Co-immunoprecipitation and immunofluorescence can probe TSPAN1-integrin interactions. This model also serves as a platform for high-content drug screening targeting tetraspanin-dependent pathways. For further information, protocols, or ordering, please contact Ascent Research.
