ZNF462 CRISPR Activation Plasmid (h): sc-412915-ACT

ZNF462 encodes a zinc finger transcription factor implicated in chromatin-dependent regulation of gene expression during development and cell fate specification. It is thought to modulate transcriptional programs by coordinating sequence-specific DNA binding with epigenetic control mechanisms that influence lineage commitment, proliferation, and differentiation. Disruption or altered dosage of ZNF462 has been associated with neurodevelopmental phenotypes and craniofacial abnormalities, supporting a role in developmental gene regulatory networks. In experimental systems, ZNF462 is studied for its impact on transcriptional control, chromatin organization, and downstream pathways governing neuronal and mesenchymal differentiation.

ZNF462 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ZNF462 expression without altering the underlying DNA sequence.

ZNF462 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ZNF462 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 ZNF462 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous ZNF462 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ZNF462 locus and enabling the study of ZNF462-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of ZNF462 pathway restoration in tumor cells with silenced or reduced ZNF462 expression.

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