iASPP CRISPR Activation Plasmid (h): sc-404703-ACT

Human PPP1R13L encodes iASPP, a regulatory protein best known as an inhibitor of p53 family transcription factors, including TP53, TP63, and TP73. By modulating p53-dependent transcriptional programs, iASPP influences apoptosis, cell-cycle control, and stress responses, integrating into pathways that shape genome integrity and cellular survival. iASPP has also been linked to cytoskeletal dynamics and cell motility through interactions that impact actin-associated signaling. Dysregulated PPP1R13L/iASPP expression has been reported in multiple cancer contexts and is investigated for roles in tumor progression, invasion, and resistance to cellular stress.

iASPP CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PPP1R13L expression without altering the underlying DNA sequence.

iASPP CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PPP1R13L 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 PPP1R13L transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous iASPP expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PPP1R13L locus and enabling the study of iASPP-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of iASPP pathway restoration in tumor cells with silenced or reduced PPP1R13L expression.

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