The GPD1L Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population targeting the GPD1L gene in the HAP1 human cell line. This heterogeneous pool of edited cells provides a loss-of-function model for studying glycerol-3-phosphate dehydrogenase 1-like protein function. The polyclonal format avoids clonal biases inherent to single-cell-derived knockouts, enabling robust functional genomic analyses. Researchers can use these cells to interrogate GPD1L-dependent metabolic and signaling networks in a defined genetic background.
HAP1 is a near-haploid human hematopoietic cell line derived from the KBM-7 chronic myeloid leukemia line. Its haploid karyotype ensures that a single CRISPR/Cas9-mediated genetic modification can effectively disrupt gene function, obviating the need for biallelic targeting. This feature, combined with stable growth and tractability, has made HAP1 a widely used platform for large-scale knockout screens, drug target discovery, and mechanistic studies. While not of cardiac origin, HAP1 cells express the core metabolic enzymes necessary to recapitulate key aspects of GPD1L biology.
GPD1L catalyzes the conversion of glycerol-3-phosphate into dihydroxyacetone phosphate, coupled to the reduction of NAD+ to NADH, thereby regulating the cytoplasmic NADH/NAD+ ratio. This redox balance critically modulates the activity of the cardiac sodium channel SCN5A, linking cellular metabolism to ion channel function and cardiac excitability. GPD1L thus acts downstream of metabolic cues such as hypoxia and NADH/NAD+ imbalance, and upstream of SCN5A-mediated sodium current. It interacts directly or indirectly with SCN5A and glycerol-3-phosphate shuttle enzymes like GPD2. Mutations disrupting this pathway are associated with Brugada syndrome and sudden infant death syndrome.
In the HAP1 background, knockout of GPD1L permits dissection of how glycerol-3-phosphate shuttle dysfunction alters the NADH/NAD+ redox state and impacts downstream effectors, including mitochondrial respiration. Although HAP1 cells lack endogenous SCN5A expression, they can be engineered to express this channel for electrophysiological assessment, providing a platform to directly measure GPD1L??s influence on sodium current. The polyclonal population reduces the risk of off-target clonal effects, offering a more representative phenotype for high-throughput screening and functional genomics studies.
Key applications include quantitative measurement of NAD+/NADH ratios using enzymatic or fluorescent assays, metabolomic profiling to map redox-related metabolites, and Seahorse metabolic flux analysis to evaluate glycolytic and oxidative phosphorylation rates. The cells are also suitable for co-immunoprecipitation to probe GPD1L?CSCN5A interactions and for drug screens targeting metabolic or ion channel modulators relevant to Brugada syndrome. For additional technical details or ordering information, please contact Ascent Research.
