KGA Lentiviral Activation Particles (h): sc-402575-LAC

Human GLS encodes glutaminase, and the kidney-type isoform KGA catalyzes mitochondrial conversion of glutamine to glutamate and ammonia, supplying carbon and nitrogen for the TCA cycle, nucleotide synthesis, and redox balance via glutathione. This enzymatic step is central to glutaminolysis and coordinates with anaplerotic metabolism, amino acid homeostasis, and cellular stress responses. Altered GLS/KGA activity is frequently studied in contexts of metabolic rewiring, including proliferative signaling, hypoxia adaptation, and oxidative stress management. Dysregulated glutamine utilization and glutamate production have been linked to mechanisms relevant to tumor metabolism, neurobiology, and immune cell activation, making GLS a common target for pathway interrogation.

KGA Lentiviral Activation Particles (h) address this need by packaging the complete synergistic activation mediator (SAM) transcriptional activation system into transduction-ready, high-titer lentiviral particles, enabling efficient GLS upregulation across a broader range of human cell types.

KGA Lentiviral Activation Particles (h) deliver all functional components of the synergistic activation mediator (SAM) system via lentiviral transduction. The system comprises three particle preparations co-transduced into target cells: one encoding catalytically inactive dCas9 (D10A and N863A mutations) fused to the VP64 transactivation domain with a blasticidin resistance gene; one encoding the MS2-p65-HSF1 fusion protein with a hygromycin resistance gene; and one encoding a target-specific 20 nt sgRNA fused to two MS2 RNA aptamers with a puromycin resistance gene. Following lentiviral transduction and genomic integration of the expression cassettes, the SAM components are stably expressed and assemble at the target locus within the proximal promoter region upstream of the GLS transcriptional start site, where VP64, p65, and HSF1 act cooperatively to recruit endogenous transcriptional machinery and drive sustained upregulation of endogenous KGA expression. The use of nuclease-inactive dCas9 avoids the introduction of double-strand DNA breaks and preserves the native GLS genomic locus and regulatory architecture.

The lentiviral format offers several practical advantages: stable genomic integration supports heritable activation across cell divisions; high-titer particle preparations eliminate the need for in-house viral production; and compatibility with primary, non-dividing, and transfection-resistant cell types expands experimental accessibility. Successful transduction can be confirmed and enriched through triple antibiotic selection using puromycin, hygromycin, and blasticidin.

For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.