I1PP2A CRISPR Activation Plasmid (m): sc-419111-ACT

Mouse Anp32a encodes I1PP2A, a nuclear acidic leucine-rich phosphoprotein that functions as an inhibitor and modulator of protein phosphatase 2A (PP2A), thereby influencing phosphorylation-dependent signaling networks. Through PP2A regulation and chromatin-associated activities, ANP32A contributes to control of cell cycle progression, DNA damage responses, and transcriptional programs linked to differentiation and stress adaptation. It has also been implicated in apoptosis regulation and broader RNA and protein homeostasis through interactions with nuclear complexes. Dysregulated ANP32A/I1PP2A activity and altered PP2A signaling are relevant to studies of oncogenic kinase pathways, neurobiology, and inflammatory or stress-related phenotypes where phosphatase balance shapes cellular outcomes.

ANP32A CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Anp32a expression without altering the underlying DNA sequence.

ANP32A CRISPR Activation Plasmid (m) 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.