EHZF CRISPR Activation Plasmid (h): sc-404324-ACT

ZNF521 encodes the human zinc finger protein EHZF, a nuclear transcriptional regulator that modulates gene expression programs linked to cell fate control, proliferation, and lineage commitment. Through its DNA-binding zinc finger domains and interactions with co-regulatory complexes, EHZF influences chromatin-dependent transcriptional networks important in hematopoietic and neural contexts. Dysregulated ZNF521 expression has been associated with altered differentiation states and aberrant growth signaling observed across several cancer-related models, making it relevant for studies of oncogenic transcriptional circuitry. As a node in broader transcription and epigenetic pathways, EHZF is frequently examined for its impact on stem-like phenotypes, developmental gene programs, and context-dependent regulatory feedback.

EHZF CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ZNF521 expression without altering the underlying DNA sequence.

EHZF CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ZNF521 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 ZNF521 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous EHZF expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ZNF521 locus and enabling the study of EHZF-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of EHZF pathway restoration in tumor cells with silenced or reduced ZNF521 expression.

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