ZNF395 CRISPR Activation Plasmid (h): sc-403829-ACT

ZNF395 encodes a zinc finger transcription factor implicated in the regulation of gene expression programs linked to cellular stress adaptation, including hypoxia-responsive transcriptional networks. It has been reported to interface with inflammatory and innate immune signaling outputs, influencing cytokine-associated pathways and broader transcriptional reprogramming. Dysregulated ZNF395 activity has been associated with altered cell state control and has been explored in the context of tumor biology and other conditions where hypoxia and inflammation shape disease-relevant phenotypes. As a nuclear DNA-binding protein, ZNF395 is a useful node for dissecting transcriptional circuitry governing proliferation, survival, and microenvironmental responses.

ZNF395 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ZNF395 expression without altering the underlying DNA sequence.

ZNF395 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ZNF395 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 ZNF395 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous ZNF395 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ZNF395 locus and enabling the study of ZNF395-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of ZNF395 pathway restoration in tumor cells with silenced or reduced ZNF395 expression.

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