Hk2 Knockout Cell Line (RAW264.7)

The choice depends on whether you are studying Hk2 (hexokinase 2)'s role as a key glycolytic enzyme in macrophage metabolic reprogramming (Warburg-like metabolism) or modeling its functions in inflammatory macrophage activation. The Knockout line is the standard tool for asking whether Hk2 is required for these processes — Hk2 is the rate-limiting first step of glycolysis (glucose → glucose-6-phosphate), and is uniquely upregulated in activated macrophages, cancer cells, and other glycolysis-dependent cells; Hk2 also has non-canonical roles in mitochondrial outer membrane association via VDAC and apoptosis regulation. Overexpression is useful for studying Hk2 gain-of-function effects. For macrophage immunology research, the EDITGENE Hk2 Knockout in RAW 264.7 is highly informative — RAW 264.7 is the gold-standard murine macrophage cell line, and Hk2 upregulation is a hallmark of LPS-induced M1 macrophage metabolic reprogramming. Other hexokinase (HK1, HK3) expression analysis aids interpretation given partial functional overlap. Rescue with wild-type or catalytically-dead Hk2 enables structure-function studies. The knockout is valuable for studying macrophage immunometabolism, Hk2-VDAC mitochondrial association, and emerging Hk2-targeted therapeutics (2-deoxyglucose, lonidamine, 3-bromopyruvate).

Primary applications: • Glycolytic flux: ¹³C-glucose tracing into glycolytic intermediates, lactate production, and glucose consumption in Hk2-null macrophages. • Macrophage immunometabolism: LPS-induced glycolytic switching (lactate production, ECAR by Seahorse) following M1 polarization in Hk2-null background. • Hk2-VDAC mitochondrial association: Hk2 mitochondrial localization and apoptosis resistance studies given Hk2's mitochondrial outer membrane association. • Hk2 inhibitor specificity: critical genetic control for 2-deoxyglucose, lonidamine, 3-bromopyruvate, and other glycolysis-targeting compounds. EDITGENE recommends this RAW 264.7-based model for researchers investigating macrophage immunometabolism, Warburg-like metabolic reprogramming, and Hk2-targeted metabolic therapy.

Yes. Hk2 rescue experiments are well-established for metabolic research: • Construct design: use a codon-modified Hk2 sequence with a small C-terminal tag (FLAG, HA). Hk2 has tandem N- and C-terminal hexokinase domains and N-terminal mitochondrial outer membrane association sequence — preserve all elements. • Catalytically-dead rescue: substrate-binding pocket mutations abolish hexokinase activity and serve as the standard specificity control. • Mitochondrial-binding-deficient rescue: N-terminal MOM-association motif mutations (e.g., 'NB' deletion) generate cytosolic Hk2 for studying VDAC-association versus catalytic functions. • Functional readout: rescue should restore glycolytic flux measured by lactate production and Seahorse ECAR. RAW 264.7-specific considerations: • RAW 264.7 is a chemically induced (Abelson murine leukemia virus-transformed) BALB/c-derived macrophage cell line — the most widely used continuous murine macrophage model for innate immunity, inflammation, and macrophage metabolism research. • Lentiviral transduction is supported with moderate efficiency. • Macrophage polarization (M1/M2) responses are preserved; characterize basal activation state before phenotypic assays.