PHF19 CRISPR Activation Plasmid (h): sc-408215-ACT

PHF19 (PHD finger protein 19) is a chromatin-associated regulator that interfaces with Polycomb repressive complex 2 (PRC2) to modulate deposition of H3K27me3 and maintain transcriptional repression programs. Through its epigenetic reader and recruitment functions, PHF19 contributes to control of cell identity, proliferation, and differentiation by shaping chromatin accessibility and gene expression states. Dysregulation of PHF19 expression or PRC2-linked chromatin pathways is implicated in oncogenic transcriptional programs and altered lineage commitment across multiple cancer contexts. As a node in Polycomb-mediated silencing, PHF19 is frequently studied for its impact on developmental gene networks, chromatin remodeling, and transcriptional plasticity.

PHF19 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PHF19 expression without altering the underlying DNA sequence.

PHF19 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PHF19 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 PHF19 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous PHF19 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PHF19 locus and enabling the study of PHF19-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of PHF19 pathway restoration in tumor cells with silenced or reduced PHF19 expression.

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