EPAS1 Knockout AC16 Cell Line

The EPAS1 Knockout AC16 Cell Line is a CRISPR/Cas9-edited human cardiomyocyte model with targeted disruption of the EPAS1 gene, encoding the hypoxia-inducible transcription factor HIF-2??. Derived from the AC16 immortalized human ventricular cardiomyocyte line, this knockout cell line enables stable loss-of-function studies of EPAS1-dependent signaling in a relevant cardiac context. By abolishing EPAS1 expression, researchers can specifically dissect HIF-2?? contributions to hypoxia-regulated transcriptional programs, avoiding crosstalk from other HIF isoforms. The product is provided for expansion and reproducible experimentation in cardiovascular and hypoxia research.

The AC16 cell line is an immortalized human adult ventricular cardiomyocyte model that maintains key morphological and biochemical features of primary cardiomyocytes, including cardiac-specific marker expression. It is widely utilized for studying cardiac physiology, drug-induced cardiotoxicity, and metabolic pathways, offering a human-relevant platform with enhanced proliferative capacity and culture convenience compared to primary cells. These attributes make AC16 particularly advantageous for long-term genetic manipulation studies and high-throughput screening, providing a controlled background for interrogating gene function in ventricular cardiomyocytes.

EPAS1 (HIF-2??) acts as a central transcriptional regulator of cellular adaptation to hypoxia. Under normoxia, prolyl hydroxylases such as EGLN1/PHD2 hydroxylate EPAS1, promoting its recognition by the VHL ubiquitin ligase and proteasomal degradation. Hypoxia inhibits hydroxylation, stabilizing EPAS1, which then translocates to the nucleus and dimerizes with ARNT (HIF-1??). The EPAS1-ARNT complex recruits CBP/p300 to activate genes with hypoxia-response elements, including VEGFA, EPO, and SLC2A1 (GLUT1). This signaling network integrates inputs from oxygen tension, growth factors (e.g., IGF-1), and cytokines (e.g., IL-6), and its dysregulation is associated with familial erythrocytosis type 4, clear cell renal cell carcinoma, and pulmonary hypertension.

In ventricular cardiomyocytes, EPAS1 mediates adaptive responses to ischemic stress through metabolic reprogramming, angiogenic signaling, and pro-survival pathways. The EPAS1 knockout in AC16 cells enables precise dissection of HIF-2??-specific functions without confounding compensation from HIF-1??. This model is instrumental for studying cardiac adaptation to hypoxia and the molecular basis of pathologies such as ischemic cardiomyopathy and right ventricular dysfunction in pulmonary hypertension. Researchers can leverage this cell line to map EPAS1-dependent transcriptional networks and evaluate their influence on cardiomyocyte viability, contractility, and metabolic homeostasis.

This cell line supports hypoxia exposure assays, Western blotting, immunofluorescence, and RT-qPCR for analyzing HIF-2?? and its targets like VEGFA and EPO. Functional studies can assess proliferation, glucose uptake, and metabolic flux under normoxia and hypoxia. The model also enables drug screening for EPAS1 modulators or VHL/PHD2 pathway inhibitors. For further inquiries, contact Ascent Research.