The GYG1 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-mediated gene-disrupted polyclonal population derived from the HAP1 human near-haploid cell line. These cells provide a loss-of-function model for glycogenin-1 (GYG1), the initiator of glycogen synthesis, allowing dissection of glycogen metabolism and insulin signaling in a genetically tractable background.
HAP1 is a chronic myeloid leukemia (CML)-derived cell line stably expressing the BCR-ABL1 fusion oncoprotein. Its near-haploid karyotype minimizes gene redundancy and simplifies knockout validation, while retaining key CML-associated signaling networks. The adherent growth and rapid doubling time make it ideal for large-scale functional assays.
GYG1 catalyzes the transfer of glucose from UDP-glucose to a tyrosine residue within its own sequence, generating an oligosaccharide primer that is extended by glycogen synthase (GYS1) to build glycogen particles. This autoglucosylation step is regulated by insulin?CINSR?CIRS1?CAKT signaling and glucose availability, with transcriptional control exerted by FOXO1 and PPARGC1A. GYG1 directly interacts with AMPK, PPP1R3C, and other glycogen metabolism components including GBE1, UGP2, and PGM1. Downstream, GYS1 activity and net glycogen accumulation rely on GYG1-mediated priming, which is also influenced by GSK3B-mediated phosphorylation.
In the HAP1 CML context, ablation of GYG1 permits examination of how cancer cells adapt their glycogen reserves under nutrient stress, BCR-ABL1-driven metabolic reprogramming, and therapeutic perturbations. The model is valuable for studying glycogen storage disease type XV, characterized by GYG1 mutations, glycogen depletion, and cardiomyopathy. Near-haploid genetics enhance the ability to identify synthetic interactions by exposing metabolic vulnerabilities not apparent in diploid cells.
This polyclonal knockout population is suitable for glycogen quantification assays, western blotting of GYG1, GYS1, phospho-AKT, and other insulin pathway nodes, glucose uptake measurements, and cell cycle or proliferation studies via flow cytometry and MTS/MTT. RNA-seq can be applied to map transcriptional adaptations. The polyclonal nature avoids clonal bias, making it appropriate for pooled screens and robust loss-of-function experiments. For further information or to place an order, please contact Ascent Research.
