PSAT1 CRISPR Activation Plasmid (h): sc-403001-ACT

Human PSAT1 encodes phosphoserine aminotransferase 1, a pyridoxal phosphate–dependent enzyme that catalyzes the conversion of 3-phosphohydroxypyruvate to 3-phosphoserine in the de novo L-serine biosynthesis pathway. By supporting serine and glycine availability, PSAT1 contributes to one-carbon metabolism, nucleotide biosynthesis, redox homeostasis, and mitochondrial function, linking central carbon metabolism to proliferative capacity. PSAT1 regulation is frequently studied in the context of ATF4-driven amino acid stress responses and integrated stress signaling, with downstream effects on anabolic growth programs. Dysregulated PSAT1 expression has been associated with metabolic reprogramming observed across multiple disease-relevant cellular models, including cancer metabolism and neurodevelopmental phenotypes.

PSAT1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PSAT1 expression without altering the underlying DNA sequence.

PSAT1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PSAT1 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 PSAT1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous PSAT1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PSAT1 locus and enabling the study of PSAT1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of PSAT1 pathway restoration in tumor cells with silenced or reduced PSAT1 expression.

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