ZNF326 CRISPR Activation Plasmid (h): sc-404968-ACT

ZNF326 encodes a zinc finger–containing nuclear protein implicated in transcriptional regulation and RNA processing, with reported roles in coordinating co-transcriptional splicing and gene expression programs that support cell-state maintenance. As a chromatin-associated factor, ZNF326 can influence promoter-proximal regulatory architecture and coupling between transcription and pre-mRNA maturation, linking it to pathways governing proliferation, differentiation, and stress-adaptive gene expression. Dysregulated ZNF326 expression or activity has been associated in the literature with altered transcriptional networks in cancer biology and other contexts where aberrant RNA metabolism contributes to disease phenotypes. These features make ZNF326 a useful target for mechanistic studies of transcription–splicing crosstalk and regulatory network rewiring.

ZNF326 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ZNF326 expression without altering the underlying DNA sequence.

ZNF326 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ZNF326 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 ZNF326 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous ZNF326 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ZNF326 locus and enabling the study of ZNF326-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of ZNF326 pathway restoration in tumor cells with silenced or reduced ZNF326 expression.

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