ATXN3 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population designed to disrupt the human ATXN3 gene in the near-haploid HAP1 cell line. This loss-of-function model enables investigation of ATXN3-dependent processes without diploid genetic complexities, offering a robust tool for studying the deubiquitinating enzyme??s role in cellular homeostasis. The polyclonal format provides a heterogeneous knockout pool suitable for population-level analyses, minimizing clonal selection artifacts.
The HAP1 host cell line originates from a male patient with chronic myeloid leukemia and exhibits a near-haploid karyotype, simplifying genetic manipulation for knockout studies. Its haploid nature ensures that single-gene disruptions reveal phenotypic consequences directly, without second-allele compensation. Widely utilized in genetic knockout research, HAP1 cells support efficient CRISPR/Cas9 editing and diverse downstream assays, serving as an ideal platform for investigating genes such as ATXN3.
ATXN3 encodes a deubiquitinating enzyme that regulates substrates involved in DNA repair, apoptosis, and protein quality control. It is activated by ATM and ATR kinases and modulates the stability of p53, PTEN, parkin, and VCP/p97. ATXN3 interacts with ubiquitin chains, polyglutamine-expanded proteins, and VCP, functioning within the ubiquitin-proteasome system, autophagy (via p62/SQSTM1 and LC3), DNA damage response, and NF-??B signaling. Through its deubiquitinase activity, ATXN3 balances protein degradation and cell survival; loss-of-function disrupts these regulatory circuits.
In HAP1 cells, ATXN3 knockout provides a defined context to model Machado-Joseph disease (spinocerebellar ataxia type 3), caused by polyglutamine expansions in ATXN3. Loss of normal ATXN3 function impairs misfolded protein clearance and sensitizes cells to proteotoxic and genotoxic stress, mirroring disease pathology. Dysregulation of p53 and NF-??B signaling also links ATXN3 to cancer biology, where it may act as a tumor suppressor or promoter. The haploid background ensures clear genotype-phenotype correlations for mechanistic studies.
These polyclonal knockout cells support diverse applications, including mechanistic studies of protein homeostasis, DNA damage responses, and neurodegeneration. Typical assays include Western blotting for ATXN3 and targets like p53 and PTEN, ubiquitination assays, proteasome activity measurements, immunofluorescence for aggregates, and apoptosis detection via Annexin V staining. They are also suited for NF-??B reporter assays and RT-qPCR for transcript ablation. This product is valuable for drug screening in polyglutamine disorders and ubiquitin-proteasome system research in cancer. For additional information, please contact Ascent Research.
