Wig-1 CRISPR Activation Plasmid (h): sc-406638-ACT

ZMAT3 (Wig-1) is a p53-inducible zinc-finger RNA-binding protein that regulates post-transcriptional gene expression by binding AU-rich elements and stabilizing or modulating the translation of select mRNAs. Through control of RNA turnover and stress-responsive transcripts, Wig-1 influences cell-cycle progression, apoptosis, and RNA surveillance programs that shape cellular responses to DNA damage and oncogenic stress. Dysregulated ZMAT3 activity has been linked to altered p53 network output and changes in tumor-associated gene expression signatures, making it relevant for mechanistic studies in cancer biology and stress signaling. Its RNA-binding functions also support investigation of mRNA stability pathways and context-dependent regulation of pro- and anti-apoptotic transcripts.

Wig-1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ZMAT3 expression without altering the underlying DNA sequence.

Wig-1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ZMAT3 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 ZMAT3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Wig-1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ZMAT3 locus and enabling the study of Wig-1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Wig-1 pathway restoration in tumor cells with silenced or reduced ZMAT3 expression.

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