ENX-2 CRISPR Activation Plasmid (h): sc-403611-ACT

Human EZH1 encodes the Polycomb-group protein ENX-2, a catalytic subunit of PRC2 that deposits the repressive histone mark H3K27me3 to regulate chromatin compaction and long-term gene silencing. ENX-2 helps control lineage-specific transcriptional programs during development and differentiation, linking epigenetic regulation to cell-cycle control and maintenance of cellular identity. As part of PRC2, EZH1 functions alongside core partners such as EED and SUZ12 to coordinate transcriptional repression across Polycomb target loci. Dysregulated PRC2 activity and altered H3K27 methylation landscapes are frequently studied in cancer biology, stem cell state transitions, and neurodevelopmental gene regulation, making EZH1 a relevant node for investigating epigenetic mechanisms.

ENX-2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous EZH1 expression without altering the underlying DNA sequence.

ENX-2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the EZH1 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 EZH1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous ENX-2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native EZH1 locus and enabling the study of ENX-2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of ENX-2 pathway restoration in tumor cells with silenced or reduced EZH1 expression.

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