The HARS1 Knockout HAP1 Polyclonal Cells product consists of a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human HAP1 cell line, engineered for loss-of-function studies of the HARS1 gene. This polyclonal knockout pool enables robust investigation of histidyl-tRNA synthetase function without clonal selection artifacts, providing a physiologically relevant model for translational and non-canonical signaling research.
HAP1 cells are a near-haploid, adherent cell line with fibroblastoid morphology, originally isolated from the male KBM-7 chronic myeloid leukemia line. The near-haploid karyotype simplifies genetic analysis and reduces the likelihood of functional compensation by wild-type alleles, making HAP1 an ideal host for CRISPR-based gene disruption studies. This background is widely used for probing gene function in cancer biology and signal transduction.
HARS1 encodes cytoplasmic histidine?CtRNA ligase, which catalyzes the ATP-dependent attachment of histidine to its cognate tRNA(His), an essential step in protein translation. Beyond its canonical role, secreted HARS1 exerts non-canonical cytokine-like activities, activating endothelial cells and contributing to inflammatory processes. Molecular interactions include eukaryotic elongation factor 1A (EEF1A), components of the multi-synthetase complex, and endothelial cell surface receptors. Upstream, HARS1 activity is regulated by amino acid availability and mTOR signaling, while downstream it enables ribosomal elongation and modulates endothelial cell activation.
In HAP1 cells, disruption of HARS1 perturbs both translational fidelity and potential non-canonical signaling axes, offering a unique platform to dissect histidyl-tRNA synthetase functions in a chronic myeloid leukemia-derived, fibroblastoid context. This model is particularly valuable for studying the mechanistic underpinnings of diseases linked to HARS1 mutations, including Usher syndrome type 3B and Charcot-Marie-Tooth disease axonal type 2W, as well as peripheral neuropathy. The near-haploid background facilitates unambiguous genotype-phenotype correlations.
Researchers can employ this knockout model for diverse applications, such as assessing translation efficiency via puromycin incorporation assays, quantifying aminoacylation activity, and probing non-canonical cytokine effects using ELISA and endothelial cell activation assays. The product is also suited for functional validation in signal transduction studies, drug screening for neurological disorders, and investigating metabolic regulation by mTOR. Standard characterization methods include Western blotting for protein expression, RT-qPCR for transcript analysis, and viability/proliferation assays. For additional information or to discuss custom projects, please contact Ascent Research.
