The DNPEP Knockout K-562 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population featuring targeted disruption of the DNPEP gene in the K-562 human chronic myelogenous leukemia cell line. This loss-of-function model provides a heterogeneous pool of cells with gene disruptions, suitable for population-based studies without clonal selection.
K-562 is an immortalized suspension cell line derived from a patient with chronic myelogenous leukemia in blast crisis. The cells harbor the BCR-ABL1 fusion gene and are Philadelphia chromosome-positive, leading to constitutive activation of proliferative pathways. Widely used as a model for leukemia and erythroid differentiation, K-562 cells grow in suspension and are amenable to high-throughput genetic manipulation and drug screening.
DNPEP encodes an aspartyl aminopeptidase that cleaves N-terminal aspartate from peptide substrates such as angiotensin II, thereby generating angiotensin III. In the renin-angiotensin system, DNPEP acts downstream of ACE and upstream of AT1 and AT2 receptors. Loss of DNPEP activity prevents angiotensin III formation, potentially altering downstream signaling through these receptors. Although its regulatory mechanisms are not fully defined, DNPEP function may be influenced by oxidative stress and hormonal cues. Disruption of this peptidase can modulate ERK phosphorylation and processes like cell proliferation and apoptosis.
In the context of K-562 leukemia cells, the renin-angiotensin system contributes to the regulation of proliferation and survival. DNPEP knockout abrogates aspartyl aminopeptidase activity, blocking the conversion of angiotensin II to angiotensin III and dysregulating downstream AT1/AT2 receptor signaling. This provides a valuable tool to study how local angiotensin metabolism influences hematopoietic cell behavior, and to investigate compensatory peptidase activities in leukemic cells.
Applications include functional dissection of DNPEP in leukemia and hematopoietic RAS signaling, screening of aminopeptidase inhibitors, and peptidase activity studies. Typical assays encompass Western blotting, RT-qPCR, angiotensin peptide quantification by ELISA/LC-MS, AT1 receptor activation assays, cell proliferation (MTS), apoptosis (Annexin V), and phospho-ERK analysis. For further details, please contact Ascent Research.
