GLDC CRISPR Activation Plasmid (h): sc-403511-ACT

Human GLDC encodes glycine decarboxylase, the P-protein component of the mitochondrial glycine cleavage system that catalyzes glycine decarboxylation and supports one-carbon unit transfer to tetrahydrofolate. Through this activity, GLDC links amino acid catabolism with folate-dependent one-carbon metabolism, influencing nucleotide biosynthesis, redox balance, and mitochondrial carbon flux. Perturbation of GLDC function disrupts glycine homeostasis and one-carbon pathways, and GLDC is implicated in inborn errors of glycine metabolism and broader metabolic dysregulation contexts. As a mitochondrial enzyme central to glycine utilization, GLDC is frequently studied for its role in cellular metabolic adaptation and regulation of biosynthetic capacity.

GLDC CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous GLDC expression without altering the underlying DNA sequence.

GLDC CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the GLDC locus in human cell lines. The system is built around a catalytically inactive Cas9 (dCas9) carrying two inactivating mutations (D10A and N863A) that eliminate nuclease activity while preserving DNA binding. This dCas9 is fused to VP64, a potent transcriptional activator, and is co-expressed with a blasticidin resistance gene for selection. The second plasmid encodes the MS2-p65-HSF1 fusion protein, a secondary activator complex that works in concert with dCas9-VP64, alongside a hygromycin resistance gene. The third plasmid encodes a target-specific 20 nt sgRNA fused to two MS2 RNA aptamers that recruit the MS2-p65-HSF1 complex to the activation site, accompanied by a puromycin resistance gene. The three plasmids are delivered at a 1:1:1 mass ratio for balanced expression of all system components.

Once assembled at the target locus, the SAM complex binds within approximately 200 bp upstream of the GLDC transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous GLDC expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native GLDC locus and enabling the study of GLDC-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of GLDC pathway restoration in tumor cells with silenced or reduced GLDC expression.

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