GPT2 Lentiviral Activation Particles (h2): sc-403739-LAC-2

Human GPT2 (glutamic pyruvate transaminase 2; mitochondrial alanine aminotransferase) encodes a PLP-dependent enzyme that catalyzes reversible transamination between alanine and α-ketoglutarate to generate pyruvate and glutamate, linking amino acid turnover to mitochondrial carbon flux. GPT2 activity interfaces with central metabolic programs including the TCA cycle, anaplerosis, and nitrogen handling, thereby influencing cellular redox balance and bioenergetic adaptation under nutrient stress. Dysregulated GPT2 expression or function has been implicated in metabolic remodeling observed in proliferative states and in neurological phenotypes where mitochondrial amino acid metabolism is perturbed. Gene editing of GPT2 supports mechanistic studies of mitochondrial metabolism, tracer-based flux analyses, and interrogation of metabolic dependencies in human cell models.

GPT2 Lentiviral Activation Particles (h2) address this need by packaging the complete synergistic activation mediator (SAM) transcriptional activation system into transduction-ready, high-titer lentiviral particles, enabling efficient GPT2 upregulation across a broader range of human cell types.

GPT2 Lentiviral Activation Particles (h2) 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 GPT2 transcriptional start site, where VP64, p65, and HSF1 act cooperatively to recruit endogenous transcriptional machinery and drive sustained upregulation of endogenous GPT2 expression. The use of nuclease-inactive dCas9 avoids the introduction of double-strand DNA breaks and preserves the native GPT2 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.