The GNPDA1 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal cell population derived from HAP1 cells, featuring heterogeneous disruption of the GNPDA1 gene. This loss-of-function model avoids clonal bias and provides a robust system for studying GNPDA1-dependent metabolic and signaling processes, suitable for pathway analysis and functional genomics applications.
HAP1 is a near-haploid human cell line originally derived from the KBM-7 chronic myeloid leukemia line from a male patient. Its near-haploid karyotype, retaining a single copy of most chromosomes, facilitates unambiguous genotype?Cphenotype correlations and is widely employed in CRISPR-based screens, drug target validation, and systematic gene perturbation studies.
GNPDA1 encodes glucosamine-6-phosphate deaminase, which catalyzes the deamination of glucosamine-6-phosphate to fructose-6-phosphate and ammonia. This reaction integrates amino sugar metabolism with glycolysis and maintains cellular pools of UDP-N-acetylglucosamine (UDP-GlcNAc), the donor substrate for O-GlcNAc transferase (OGT)-mediated O-GlcNAcylation and N-glycan biosynthesis. GNPDA1 is regulated by insulin/IGF-1 signaling, HIF-1A, OGT, glucose availability, and AMPK. Its disruption impairs downstream O-GlcNAc cycling, affecting mTOR signaling and protein O-GlcNAcylation substrates. The enzyme interacts with hexokinase, phosphoglucose isomerase, and glutamine-fructose-6-phosphate transaminase (GFPT1/2), positioning it at a critical node between energy sensing and hexosamine pathway flux.
In the HAP1 genetic background, loss of GNPDA1 produces a penetrant phenotype due to near-haploidy, making it ideal for dissecting hexosamine pathway contributions to cancer metabolism, type 2 diabetes, and neurodegenerative diseases. Depletion of UDP-GlcNAc globally reduces O-GlcNAcylation, altering nutrient sensing, transcriptional regulation, and stress responses, and allowing systematic investigation of these processes under defined genetic perturbation.
Applications include Western blotting with O-GlcNAc-specific antibodies, RT-qPCR for GNPDA1 expression, LC-MS/MS-based metabolite profiling of hexosamine intermediates, glucose uptake and proliferation assays, and Click chemistry-based detection of O-GlcNAcylated proteins. The polyclonal format supports pooled CRISPR screens and metabolic drug target validation. For further technical inquiries, please contact Ascent Research.
