Zic3 CRISPR Activation Plasmid (h): sc-407048-ACT

ZIC3 encodes the zinc finger transcription factor Zic3, a nuclear DNA-binding protein that regulates early embryonic patterning and left–right axis specification. Zic3 modulates gene regulatory networks involved in mesendoderm formation, neural development, and cilia-dependent signaling that integrates with pathways such as NODAL/SMAD and WNT during morphogenesis. Dysregulated ZIC3 expression or function is associated with laterality defects and congenital malformations, making it a useful node for studying developmental transcriptional programs. In human cell models, ZIC3 provides a tractable readout for investigating lineage specification, chromatin-state control of developmental loci, and transcription factor–driven network rewiring.

Zic3 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ZIC3 expression without altering the underlying DNA sequence.

Zic3 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ZIC3 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 ZIC3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Zic3 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ZIC3 locus and enabling the study of Zic3-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Zic3 pathway restoration in tumor cells with silenced or reduced ZIC3 expression.

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