ACE2 Knockout NCI-H1299 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population that disrupts the ACE2 gene (angiotensin-converting enzyme 2) in the human non-small cell lung carcinoma NCI-H1299 cell line. This polyclonal pool results from targeted gene disruption without single-cell clonal expansion, providing a heterogeneous knockout model suitable for population-level functional analyses. The loss of ACE2 function allows researchers to study its roles in signaling, viral entry, and cancer biology.
The NCI-H1299 cell line was derived from a lymph node metastasis of lung adenocarcinoma and serves as a widely used epithelial model for non-small cell lung carcinoma. These cells display anchorage-independent growth and tumorigenic properties, making them valuable for dissecting mechanisms of oncogenesis, metastasis, and therapeutic response. Engineering ACE2 deficiency in this background permits investigation of the intersection between the renin-angiotensin system and lung cancer.
ACE2 is a carboxypeptidase that converts angiotensin II to angiotensin-(1-7), a peptide that activates the MAS1 receptor to counterbalance the angiotensin II/AGTR1 (AT1R) axis, promoting vasodilation, anti-inflammatory, and anti-fibrotic effects. ACE2 transcription is regulated by interferons (IFN-??, IFN-??) and transcription factors including HNF1A, GATA4, and FOXA2. Downstream, angiotensin-(1-7) stimulates nitric oxide production and phosphorylates Akt and ERK1/2. ACE2 also serves as the receptor for SARS-CoV-2 spike protein, facilitated by interaction with SLC6A19 (B0AT1). Knockout ablates both enzymatic and receptor functions.
Knockout of ACE2 in NCI-H1299 cells is expected to increase angiotensin II signaling through AT1R, potentially enhancing pro-inflammatory and proliferative pathways relevant to lung cancer biology. This model enables examination of how ACE2 deficiency impacts tumor cell behavior, including proliferation and migration. Additionally, the absence of ACE2 renders cells resistant to SARS-CoV-2 entry, establishing a controlled system for viral host factor studies and for distinguishing ACE2-dependent from independent mechanisms.
Applications include SARS-CoV-2 pseudovirus entry assays, angiotensin II cleavage activity measurements via ELISA for angiotensin-(1-7), and RNA-seq to define transcriptomic consequences of ACE2 loss. Standard validation assays such as western blotting and RT-qPCR confirm knockout, while functional rescue experiments can re-introduce ACE2 to restore phenotypes. The polyclonal pool is also suitable for drug screening targeting ACE2?Cspike interactions and for exploring RAS contributions to lung cancer. For technical support, contact Ascent Research.
