ZNF224 CRISPR Activation Plasmid (h): sc-407529-ACT

ZNF224 encodes a KRAB domain–containing C2H2 zinc finger transcription factor that regulates gene expression through sequence-specific DNA binding and recruitment of corepressor complexes such as KAP1/TRIM28, shaping chromatin state and transcriptional output. It participates in transcriptional repression programs linked to epigenetic control, cell-cycle regulation, stress responses, and differentiation, with context-dependent effects on downstream target genes. Altered ZNF224 activity has been associated with dysregulated transcriptional networks observed across multiple disease settings, including cancer-related pathways and inflammatory signaling. Because ZNF224 can modulate broad gene regulatory circuits, it is frequently studied as a node connecting chromatin regulation to phenotype.

ZNF224 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ZNF224 expression without altering the underlying DNA sequence.

ZNF224 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ZNF224 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 ZNF224 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous ZNF224 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ZNF224 locus and enabling the study of ZNF224-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of ZNF224 pathway restoration in tumor cells with silenced or reduced ZNF224 expression.

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