ITPase CRISPR Activation Plasmid (m): sc-421191-ACT

Mouse Itpa encodes inosine triphosphate pyrophosphatase (ITPase), a cytosolic “housekeeping” enzyme that hydrolyzes noncanonical purine triphosphates such as ITP and dITP to prevent their accumulation. By sanitizing the nucleotide pool, ITPase supports replication fidelity, DNA/RNA integrity, and genome maintenance processes that are tightly linked to cellular stress responses. Disruption of nucleotide pool homeostasis can increase misincorporation events and elevate DNA damage signaling, connecting ITPA function to pathways influencing mutagenesis and inflammatory signaling cascades. Altered ITPA activity is therefore relevant to models of genomic instability and metabolism-associated phenotypes in mouse systems.

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

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

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