PCGF1 CRISPR Activation Plasmid (h): sc-407339-ACT

PCGF1 (polycomb group ring finger 1) encodes a RING finger protein that functions as a core component of Polycomb repressive complex 1 (PRC1)-related assemblies, contributing to ubiquitination of histone H2A and maintenance of transcriptional silencing. Through epigenetic regulation, PCGF1 helps control chromatin compaction, cell-cycle and differentiation programs, and lineage-specific gene expression, with prominent roles in stem and progenitor cell states. PCGF1 interfaces with broader Polycomb/PRC networks and cooperates with other chromatin regulators to modulate developmental pathways and genome stability. Dysregulated Polycomb activity, including altered PCGF1-associated repression, is frequently studied in the context of oncogenic transcriptional programs, aberrant differentiation, and other diseases driven by epigenetic imbalance.

PCGF1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PCGF1 expression without altering the underlying DNA sequence.

PCGF1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PCGF1 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 PCGF1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous PCGF1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PCGF1 locus and enabling the study of PCGF1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of PCGF1 pathway restoration in tumor cells with silenced or reduced PCGF1 expression.

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