AICAR CRISPR Activation Plasmid (h): sc-404771-ACT

Human ATIC encodes the bifunctional enzyme 5-aminoimidazole-4-carboxamide ribonucleotide formyltransferase/IMP cyclohydrolase (AICAR), which catalyzes the final two steps of de novo purine biosynthesis leading to inosine monophosphate production. Through its role in one-carbon metabolism–dependent formyl transfer and nucleotide pool maintenance, ATIC supports DNA/RNA synthesis, cell-cycle progression, and metabolic homeostasis. Perturbation of ATIC activity can impact proliferative capacity and replication stress responses, linking purine pathway dysregulation to cancer cell metabolism and broader metabolic vulnerabilities. Inherited defects in this pathway have been associated with neurodevelopmental and metabolic phenotypes, underscoring the importance of tight control over purine synthesis.

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

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

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