IMPDH2 CRISPR Activation Plasmid (h): sc-410835-ACT

Human IMPDH2 encodes inosine monophosphate dehydrogenase 2, a rate-limiting enzyme in de novo guanine nucleotide biosynthesis that converts IMP to XMP and helps sustain cellular GTP pools. By controlling nucleotide availability, IMPDH2 supports DNA/RNA synthesis, ribosome biogenesis, and proliferation, linking it to metabolic remodeling programs that accompany immune activation and oncogenic growth. IMPDH2 activity intersects with purine metabolism and broader nucleotide homeostasis pathways that influence replication stress responses and transcriptional capacity. Altered expression or dependency on IMPDH2 has been reported across cancers and in contexts of immune and inflammatory cell expansion, making it a useful node for studying metabolic regulation of cell state.

IMPDH2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous IMPDH2 expression without altering the underlying DNA sequence.

IMPDH2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the IMPDH2 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 IMPDH2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous IMPDH2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native IMPDH2 locus and enabling the study of IMPDH2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of IMPDH2 pathway restoration in tumor cells with silenced or reduced IMPDH2 expression.

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