The ANKRD13A Knockout Jurkat Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population with disrupted ANKRD13A gene. This heterogeneous pool of Jurkat T lymphocytes carries diverse loss-of-function mutations, enabling unbiased interrogation of ANKRD13A functions. ANKRD13A acts as an adaptor in the ubiquitin-proteasome system, facilitating degradation of ubiquitinated substrates via ERAD and endosomal sorting. These cells provide a model to study protein quality control, receptor trafficking, and signaling attenuation.
Jurkat cells, an immortalized human CD4+ T lymphocyte line from acute lymphoblastic leukemia, are a standard model for T cell signaling, apoptosis, and leukemia. They express robust ubiquitin-proteasome and endocytic machinery, making them ideal for dissecting protein degradation. Their proliferation and defined background support reproducible biochemical, imaging, and flow cytometric analyses of protein turnover and signaling.
ANKRD13A is a ubiquitin-binding adaptor that links ubiquitinated cargo to the degradation machinery. It interacts with the SYVN1/HRD1 E3 ligase complex, VCP/p97, and the 26S proteasome to mediate ERAD of misfolded proteins. Upon EGF stimulation, ANKRD13A is recruited to ubiquitinated EGFR, directing it to lysosomal degradation and attenuating signaling via Grb2 and Cbl. Upstream, ER stress and UPR sensors (IRE1??, PERK, ATF6) upregulate ANKRD13A. Thus, ANKRD13A integrates UPR, ubiquitin-dependent proteolysis, and growth factor receptor downregulation, involving Derlin-1, Sel1L, and the HRD1 complex.
In Jurkat cells, ANKRD13A knockout offers a relevant system to study ubiquitin-dependent degradation in T cell homeostasis and leukemia. Moderate EGFR expression and active TCR signaling permit exploration of crosstalk between protein quality control and immune receptor pathways. Loss of ANKRD13A may impair misfolded protein clearance and alter receptor downregulation kinetics, providing insights into proteotoxic stress relevant to cancer and protein aggregation disorders.
These polyclonal knockout cells suit diverse assays: Western blotting for ubiquitinated proteins/EGFR and cycloheximide chases to assess protein half-life; co-IP with SYVN1 or VCP/p97 to validate interactions; flow cytometry for surface EGFR; ER stress induction (tunicamycin/thapsigargin) with apoptosis assays and proteasome activity measurements. Applications include ERAD mechanism studies, EGFR degradation analysis, protein quality control research, and drug screening for ER stress-related diseases. For additional information or custom requests, please contact Ascent Research.
