The ACE2 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population designed to disrupt the human ACE2 gene in the HAP1 host cell line. This loss-of-function model provides a genetically defined pool of cells carrying various disruptions at the target locus, enabling rapid phenotypic screening without the need for single-cell cloning. The polyclonal format maintains host genetic stability while delivering robust target-gene ablation for functional studies.
HAP1 is a near-haploid human cell line derived from the chronic myeloid leukemia cell line KBM-7, featuring a near-haploid karyotype and p53 deficiency. These properties make it a favored model for functional genomics and genetic screens, as the simplified genome reduces functional redundancy and facilitates unambiguous genotype-phenotype correlations. HAP1 cells grow in suspension and have been extensively used in CRISPR knockout libraries and drug-target validation.
ACE2 encodes a transmembrane carboxypeptidase that converts angiotensin II to angiotensin-(1-7), which acts on the Mas receptor to counterbalance vasoconstriction. ACE2 is also the primary receptor for SARS-CoV-2, where the viral spike protein binds to ACE2 and is primed by TMPRSS2 for membrane fusion. Additionally, ADAM17-mediated shedding releases soluble ACE2, modulating its activity. Downstream, ACE2 influences NF-??B and MAPK pathways and interacts with integrins and AT1R. Thus, ACE2 links the renin-angiotensin system with viral entry mechanisms.
Knocking out ACE2 in the HAP1 background eliminates the dominant cellular entry portal for SARS-CoV-2, rendering cells resistant to spike-mediated infection. The p53-deficient status avoids confounding pro-apoptotic signals during viral challenge experiments. Moreover, the near-haploid genome ensures that the ACE2 disruption directly accounts for observed phenotypes, such as loss of angiotensin II conversion or blocked pseudovirus uptake, making the model ideal for high-throughput screening of entry inhibitors or dissection of ACE2-dependent signaling.
Researchers can employ these cells in SARS-CoV-2 pseudovirus entry assays, angiotensin II conversion assays, and molecular analyses including Western blotting, RT-qPCR, immunofluorescence, and flow cytometry. Applications span virus-host interaction studies, cardiovascular research, and drug screening for ACE2-targeted interventions. This polyclonal knockout population is a versatile tool for COVID-19 biology and beyond. For further technical details, please contact Ascent Research.
