The ACE Knockout HAP1 Polyclonal Cells product comprises a CRISPR/Cas9-edited polyclonal knockout cell population in which the ACE (Angiotensin-Converting Enzyme) gene has been disrupted using CRISPR/Cas9 technology. These polyclonal cells are derived from the HAP1 parental line and feature a heterogeneous knockout of the ACE locus, enabling loss-of-function studies without the selection of a single clonal isolate. The knockout population provides a flexible model for investigating ACE-dependent signaling cascades, enzyme activity, and downstream physiological effects in a near-haploid genetic background.
The host cell line, HAP1, is a near-haploid human cell line originally derived from the KBM-7 chronic myeloid leukemia line. HAP1 cells display a predominantly haploid karyotype, which simplifies gene targeting and facilitates the generation of homogeneous knockout populations. This cell line is widely employed in functional genomic screens, protein?Cprotein interaction studies, and pharmacological assays due to its genetic tractability and human origin. The haploid state minimizes complications from diploidy-dependent gene compensation, making HAP1 an excellent platform for dissecting monogenic contributions to signaling pathways.
ACE is a membrane-bound zinc metallopeptidase pivotal in the renin-angiotensin system (RAS). It converts angiotensin I to angiotensin II and degrades bradykinin. Upstream regulators include the glucocorticoid receptor, angiotensin II, bradykinin, hypoxia-inducible factor 1-alpha (HIF1A), and thyroid hormone receptors. Downstream, angiotensin II acts through the angiotensin II receptor type 1 (AGTR1) to promote aldosterone secretion and vasopressin release, while bradykinin signals via the bradykinin receptor B2. ACE interacts with angiotensin I, renin, angiotensinogen, and ACE inhibitors such as captopril. Thus, ACE coordinates vascular tone and fluid balance at the intersection of the RAS and kallikrein?Ckinin system.
Disruption of ACE in the HAP1 polyclonal knockout population shifts the balance from vasoconstriction to vasodilation. Reduced angiotensin II lowers AGTR1 signaling and aldosterone/vasopressin output, while accumulating bradykinin enhances B2 receptor-mediated nitric oxide release. This mimics ACE inhibitor pharmacology, making the cells a valuable model for RAS research, endothelial biology, and electrolyte homeostasis. The HAP1 genetic tractability enables high-throughput screening approaches.
Applications include RAS pathway dissection, hypertension and cardiovascular disease modeling, ACE inhibitor screening, and bradykinin metabolism studies. Typical assays are ACE activity measurements, angiotensin II and bradykinin ELISAs, Western blotting, RT-qPCR, immunofluorescence, and cell migration or proliferation assays. The model also aids research on diabetic nephropathy and myocardial infarction. For additional information, contact Ascent Research.
