ZKSCAN3 CRISPR Activation Plasmid (h): sc-416370-ACT

ZKSCAN3 (zinc finger with KRAB and SCAN domains 3) is a nuclear DNA-binding transcription factor that integrates SCAN-mediated protein interactions with KRAB-dependent transcriptional regulation. It modulates gene expression programs linked to cell proliferation, differentiation, and cellular stress responses, and has been implicated in regulation of autophagy- and lysosome-associated pathways. Altered ZKSCAN3 activity has been reported in multiple disease contexts, including cancers where dysregulated transcriptional networks contribute to invasive growth and metabolic adaptation. As a node in transcriptional control, ZKSCAN3 is a useful target for dissecting upstream signaling inputs and downstream gene signatures relevant to oncogenic and stress-response biology.

ZKSCAN3 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ZKSCAN3 expression without altering the underlying DNA sequence.

ZKSCAN3 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ZKSCAN3 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 ZKSCAN3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous ZKSCAN3 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ZKSCAN3 locus and enabling the study of ZKSCAN3-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of ZKSCAN3 pathway restoration in tumor cells with silenced or reduced ZKSCAN3 expression.

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