The HACE1 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal cell population with targeted disruption of the HACE1 gene in the HAP1 human near-haploid fibroblast-like cell line. This product provides a heterogeneous knockout model suitable for pooled functional genomics screens and robust loss-of-function studies, avoiding clonal artifacts while ensuring perturbation of HACE1 function across the population.
HAP1 cells, derived from the chronic myeloid leukemia line KBM-7, possess a near-haploid karyotype that eliminates allele redundancy, enabling unambiguous phenotype-genotype correlations. Widely adopted for genetic screening, this cell line maintains stable growth and retains signaling pathways relevant to cancer cell biology. Its haploid nature ensures penetrant phenotypes upon gene disruption, making it an ideal host for knockout studies.
HACE1 is an E3 ubiquitin ligase that targets active Rac1 for ubiquitination and degradation, thereby suppressing Rac1-dependent NADPH oxidase-mediated ROS production. This regulation connects oxidative stress to autophagy: HACE1 interacts with p62/SQSTM1 and optineurin, and its expression is induced by hypoxia and oxidative stress. Downstream, loss of HACE1 leads to Rac1 accumulation, elevated ROS, and impaired autophagy initiation, disrupting critical tumor-suppressive and redox-balance mechanisms.
In the HAP1 polyclonal knockout context, HACE1 disruption results in sustained Rac1 activity and increased ROS, permitting direct investigation of HACE1-deficiency phenotypes. This model is relevant to cancers such as Wilms tumor and neuroblastoma, where HACE1 loss contributes to oncogenesis. The near-haploid background ensures clear readouts of HACE1??s roles in proliferation, death, and migration without wild-type allele interference.
The cells are suitable for assays including Rac1 ubiquitination analysis, ROS quantification, autophagy flux measurement (LC3 turnover), and phenotypic studies of proliferation, colony formation, and apoptosis. Western blotting and immunofluorescence for HACE1, Rac1, and LC3 puncta are readily applicable. Researchers in cancer biology, oxidative stress, ubiquitin signaling, and neurodegeneration will find this a versatile tool for target validation and mechanistic dissection. For additional information, please contact Ascent Research.
