The ATE1 Knockout HAP1 Polyclonal Cells are a polyclonal population of HAP1 cells with CRISPR/Cas9-mediated disruption of the ATE1 gene, providing a loss-of-function model for studying arginyltransferase 1 (ATE1)-dependent processes. The polyclonal nature ensures diverse knockout alleles, minimizing clonal bias and enabling robust population-level analyses of arginylation and the N-end rule pathway.
HAP1 cells are near-haploid chronic myeloid leukemia cells derived from the KBM-7 line, widely used for functional genomics and knockout screens due to efficient gene targeting and the absence of a second functional allele. Their hematopoietic origin offers a relevant context for investigating protein homeostasis and signaling pathways pertinent to leukemia and other disorders.
ATE1 encodes an arginyltransferase that mediates N-terminal arginylation of proteins, a key step in the N-end rule degradation pathway. Following arginine addition, substrates are recognized by the UBR1 E3 ligase and targeted to the proteasome. ATE1 expression is induced by oxidative stress and hypoxia via ATF4, and it modifies downstream factors including RGS4, RGS5, beta-actin, calreticulin, and alpha-synuclein. ATE1 forms functional complexes with UBR1, proteasomal subunits, arginyl-tRNA synthetase, and HSP70, thereby governing protein turnover, stress granule dynamics, and cytoskeletal organization.
In the near-haploid HAP1 background, ATE1 disruption eliminates arginylation activity, stabilizing N-end rule substrates and enabling clear dissection of arginylation-dependent phenotypes. The polyclonal knockout pool avoids clonal artifacts and supports population-based assays such as arginylation activity measurements, protein stability analysis, and stress sensitivity testing. This model is particularly useful for studying the interplay between arginylation and apoptosis, autophagy, and stress responses.
Applications include investigation of N-end rule pathway mechanisms, identification of novel ATE1 substrates, and functional studies of arginylation in cancer and neurodegeneration. Compatible assays include Sanger sequencing and western blotting for knockout verification, in vitro arginylation assays, Ub-R-GFP reporter stability assays, co-immunoprecipitation, and oxidative stress or apoptosis assays. The cells also facilitate drug target validation for proteasome-related pathways. For further information, please contact Ascent Research.
