PML CRISPR Activation Plasmid (h): sc-400145-ACT

Human PML encodes promyelocytic leukemia protein, a key organizer of PML nuclear bodies that coordinate transcriptional regulation, DNA damage responses, apoptosis, and intrinsic antiviral defense. PML influences post-translational SUMOylation networks and interfaces with pathways including p53 signaling, interferon-stimulated gene programs, and chromatin remodeling to shape cell fate decisions under stress. Disruption or altered regulation of PML nuclear body dynamics has been linked to oncogenic transformation and defective genome surveillance, with prominent relevance in hematologic malignancy biology and broader cancer models. As a scaffold protein with context-dependent effects, PML provides a tractable node for dissecting nuclear compartmentalization and stress-response circuitry in human cells.

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

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

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