ZNF282 CRISPR Activation Plasmid (h): sc-409149-ACT

ZNF282 encodes a zinc finger transcription factor implicated in sequence-specific DNA binding and regulation of gene expression programs that influence cell state and differentiation. As a nuclear regulatory protein, ZNF282 is associated with transcriptional control processes coordinated by RNA polymerase II and chromatin-associated cofactors, supporting context-dependent activation or repression of downstream targets. Altered ZNF282 expression has been reported in transcriptomic and functional studies across multiple cancer contexts, consistent with roles in proliferation, invasion-associated gene networks, and cellular stress responses. These features make ZNF282 a useful node for interrogating transcriptional circuitry, epigenetic regulation, and pathway rewiring in human cells.

ZNF282 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ZNF282 expression without altering the underlying DNA sequence.

ZNF282 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ZNF282 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 ZNF282 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous ZNF282 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ZNF282 locus and enabling the study of ZNF282-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of ZNF282 pathway restoration in tumor cells with silenced or reduced ZNF282 expression.

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