Zic2 CRISPR Activation Plasmid (h): sc-404720-ACT

ZIC2 encodes the zinc finger transcription factor Zic2, a key regulator of early embryonic patterning and neurodevelopmental gene programs. Zic2 binds cis-regulatory elements to control transcriptional networks involved in neural tube formation, forebrain specification, and cell fate determination, intersecting with pathways that shape morphogen responses and chromatin-state transitions. Dysregulated ZIC2 expression or function has been associated with developmental disorders including holoprosencephaly, and altered transcriptional circuitry involving ZIC2 has been reported across multiple cancer contexts. As a DNA-binding regulator, Zic2 provides a tractable node for probing gene regulatory networks, lineage specification, and transcription factor–driven phenotypes in human cell models.

Zic2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ZIC2 expression without altering the underlying DNA sequence.

Zic2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ZIC2 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 ZIC2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Zic2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ZIC2 locus and enabling the study of Zic2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Zic2 pathway restoration in tumor cells with silenced or reduced ZIC2 expression.

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