The AUH Knockout HAP1 Polyclonal Cells product consists of a CRISPR/Cas9-edited polyclonal knockout cell population derived from HAP1 cells, featuring disruption of the AUH gene (Homo sapiens). This polyclonal format provides a heterogeneous loss-of-function model suitable for studying the dual roles of AUH in post-transcriptional regulation and leucine catabolism. The gene-edited population is obtained through CRISPR/Cas9-mediated targeting, yielding a mixed population that captures diverse editing outcomes and supports robust functional analyses.
HAP1 cells are a near-haploid human cell line originally derived from the KBM-7 chronic myeloid leukemia cell line, exhibiting an adherent fibroblast-like morphology. Their haploid genomic content makes them particularly advantageous for genetic screens, loss-of-function studies, and functional genomics applications. In cancer biology and signaling research, HAP1 cells provide a simplified genetic background that facilitates the interrogation of gene function without the complexity of diploid compensation, making them an ideal host for generating knockout models.
AUH encodes a bifunctional protein that acts as an AU-rich element (ARE)-binding protein regulating mRNA stability and as a methylglutaconyl-CoA hydratase in the mitochondrial leucine degradation pathway. In leucine catabolism, AUH catalyzes the conversion of 3-methylglutaconyl-CoA to 3-hydroxy-3-methylglutaryl-CoA, functioning downstream of the branched-chain ketoacid dehydrogenase complex (including BCKDHA, BCKDHB, DBT) and upstream of HMGCL. It binds AREs in mRNAs such as TNF and IL-6, thereby modulating their turnover. AUH activity is influenced by leucine abundance and metabolic transcription factors, and it associates with other RNA-binding proteins and mitochondrial import receptors. Consequently, AUH links metabolic state to post-transcriptional gene regulation, impacting inflammatory and growth-related transcripts.
In the context of HAP1 cells, disruption of AUH creates a valuable model for dissecting the intersection of mitochondrial metabolism and RNA biology. The near-haploid background accentuates loss-of-function phenotypes, facilitating clear readouts in metabolic assays and RNA regulatory studies. This knockout model permits investigation of how AUH deficiency leads to accumulation of 3-methylglutaconyl-CoA and dysregulation of ARE-containing mRNAs, mimicking aspects of 3-methylglutaconic aciduria type I, a neurometabolic disorder. The cancer-cell derivation of HAP1 further enables exploration of metabolic vulnerabilities and altered RNA regulatory networks that may contribute to oncogenesis or neurodegeneration.
Researchers can employ this AUH polyclonal knockout cell population in a broad range of experimental applications, including metabolic flux analysis using labeled leucine to trace pathway intermediates, RNA immunoprecipitation to assess AUH target engagement, mitochondrial respiration assays to evaluate oxidative function, and cell viability screens to identify synthetic lethal interactions. The model also supports CRISPR negative selection screens and studies of post-transcriptional control of inflammation. Combined with Western blotting and RT-qPCR for confirmation of protein and transcript levels, these cells provide a versatile platform for basic and translational research. For further information, please contact Ascent Research.
