OAZ CRISPR Activation Plasmid (m): sc-430124-ACT

Mouse Zfp423 encodes the multi–zinc finger transcriptional regulator OAZ, a DNA-binding protein that integrates developmental signaling inputs to control lineage specification and differentiation programs. OAZ participates in transcriptional networks linked to BMP/SMAD signaling and interacts with cofactors that shape neuronal and adipocyte gene expression, influencing cell fate decisions and maturation. In the nervous system, Zfp423 is implicated in cerebellar development and neuronal patterning, while in metabolic tissues it contributes to adipogenesis and energy homeostasis. Dysregulation of Zfp423-associated transcriptional programs is therefore relevant to models of neurodevelopmental defects and metabolic phenotypes where altered differentiation and signaling cross-talk are key mechanisms.

OAZ CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Zfp423 expression without altering the underlying DNA sequence.

OAZ CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Zfp423 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 Zfp423 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous OAZ expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Zfp423 locus and enabling the study of OAZ-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of OAZ pathway restoration in tumor cells with silenced or reduced Zfp423 expression.

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