The ALDOC Knockout HEK293T Polyclonal Cells product is a heterogeneous population of HEK293T cells with CRISPR/Cas9-mediated disruption of the ALDOC gene, creating a loss-of-function model of fructose-bisphosphate aldolase C. This polyclonal knockout population, generated without single-cell cloning, maintains genetic diversity and enables robust loss-of-function studies in metabolic and signaling research.
The host cell line, HEK293T, is a human embryonic kidney epithelial cell derivative stably expressing the SV40 large T-antigen, which permits episomal replication and high transfection efficiency. These cells are widely used due to their reliable growth, ease of manipulation, and permissiveness to viral transduction and plasmid delivery, providing a physiologically relevant platform for studying metabolic enzymes such as ALDOC.
ALDOC encodes fructose-bisphosphate aldolase C, which reversibly cleaves fructose-1,6-bisphosphate into glyceraldehyde 3-phosphate (G3P) and dihydroxyacetone phosphate (DHAP) during glycolysis and gluconeogenesis. This reaction is a critical node in carbon metabolism, regulated by upstream factors including HIF-1??, c-Myc, insulin, and glucagon. The resulting G3P and DHAP drive ATP and pyruvate production, influencing lactate output and energy homeostasis. ALDOC interacts with actin filaments, ??-tubulin, V-ATPase, GAPDH, and phosphofructokinase, thereby linking glycolysis to cytoskeletal organization and pH regulation, and it associates with HIF-1?? within the hypoxia signaling pathway.
In the HEK293T context, characterized by high aerobic glycolysis akin to the Warburg effect, ALDOC disruption reduces glycolytic flux, leading to diminished pools of G3P, DHAP, ATP, and pyruvate, and impairing biosynthetic precursor supply. This effect disrupts metabolic reprogramming-associated phenotypes and may disturb glycolytic enzyme?Ccytoskeleton interactions, making these polyclonal knockout cells valuable for dissecting aldolase C??s integrative roles in metabolism, structure, and signaling.
Applications include Seahorse metabolic flux analysis, glycolytic rate and lactate quantification, ATP assays, and RNA-seq transcriptomics, supporting studies on HIF-1??-driven adaptation, c-Myc-dependent glycolysis, and aldolase isozyme interplay in cancer and neurology. These cells are suitable for drug screens targeting glycolytic vulnerabilities, hypoxia response research, and fructose metabolism investigations. Standard validation uses western blotting, RT-qPCR, and functional assays such as proliferation and apoptosis. For further information, contact Ascent Research.
