I1PP2A CRISPR Activation Plasmid (h): sc-402988-ACT

ANP32A encodes I1PP2A, an acidic leucine-rich nuclear phosphoprotein that functions as an inhibitor of protein phosphatase 2A (PP2A), shaping phosphorylation-dependent signaling and chromatin-associated processes. By modulating PP2A activity, I1PP2A influences cell-cycle progression, DNA damage responses, transcriptional programs, and stress-adaptive signaling networks. ANP32A has been linked to regulation of apoptosis and innate immune responses and is frequently studied in contexts where aberrant phosphatase control alters cellular homeostasis. Dysregulated ANP32A/I1PP2A expression or function has been associated with tumor biology, neurodegenerative mechanisms, and inflammatory signaling, supporting its relevance in pathway-centric disease models.

ANP32A CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ANP32A expression without altering the underlying DNA sequence.

ANP32A CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ANP32A 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 ANP32A transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous ANP32A expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ANP32A locus and enabling the study of ANP32A-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of ANP32A pathway restoration in tumor cells with silenced or reduced ANP32A expression.

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