This product consists of CRISPR/Cas9-edited polyclonal knockout A-549 cells, engineered for targeted disruption of the AKAP1 gene. The polyclonal population contains a heterogeneous mixture of gene-edited alleles, eliminating AKAP1 expression without introducing clonal biases. This loss-of-function model enables robust investigation of AKAP1-dependent signaling in a human lung adenocarcinoma background.
The A-549 cell line is derived from human lung carcinoma tissue and serves as a canonical model for lung adenocarcinoma. These epithelial cells retain key features of alveolar type II pneumocytes and commonly harbor KRAS mutations, making them valuable for studying oncogenic signaling and mitochondrial adaptations. The adherent, fast-growing nature of A-549 supports a wide array of proliferative, metabolic, and imaging-based assays. In this context, AKAP1 knockout provides a relevant platform to explore mitochondrial scaffolding functions in lung cancer.
AKAP1 functions as a mitochondrial scaffold, anchoring PKA holoenzyme via regulatory subunits RI/RII to the outer membrane. This positions PKA to respond to cAMP elevations, integrating inputs from reactive oxygen species and mitochondrial stress. Activated PKA then phosphorylates Drp1 at Ser616, promoting mitochondrial fission, and BAD at Ser155, inhibiting apoptosis. AKAP1 also interacts with PP1 and additional adaptors, forming a signaling node that governs mitochondrial dynamics and cell survival. Gene disruption uncouples PKA from its substrates, dampening cAMP-driven phosphorylation and tilting the balance toward mitochondrial dysfunction and altered apoptosis.
In A-549 lung adenocarcinoma cells, loss of AKAP1 disrupts mitochondrial PKA signaling likely influencing cancer-relevant phenotypes such as metabolic reprogramming, apoptosis resistance, and mitochondrial fragmentation. This model permits dissection of how the cAMP-PKA-AKAP1 axis sustains mitochondrial homeostasis in a tumor environment where mitochondrial dynamics are often co-opted for survival. Researchers can thus investigate whether AKAP1 deletion sensitizes cells to metabolic stress or apoptotic stimuli, potentially identifying mitochondrial vulnerabilities in lung cancer.
The AKAP1 knockout polyclonal cells are suited for diverse experimental approaches. Western blotting can assess phosphorylation of Drp1 and BAD, while MitoTracker immunofluorescence reveals mitochondrial network morphology. Flow cytometry with annexin V probes apoptosis, and Seahorse metabolic flux analysis uncovers shifts in oxidative phosphorylation and glycolysis. Co-immunoprecipitation confirms disrupted PKA-AKAP1 complexes. Cell viability assays under metabolic or chemotherapeutic stress evaluate functional outcomes. Collectively, these tools support advanced studies on mitochondrial signaling, cAMP dynamics, and cancer cell fitness. For inquiries, please contact Ascent Research.
