ZFHX4 CRISPR Activation Plasmid (m): sc-429838-ACT

Zfhx4 encodes the mouse zinc finger homeobox protein ZFHX4, a large DNA-binding transcription factor implicated in long-range transcriptional control during development. ZFHX4 is associated with regulation of lineage specification and neuronal differentiation programs, linking it to chromatin state modulation and coordinated control of gene expression networks. Altered ZFHX4 expression or regulatory disruption has been connected to neurodevelopmental phenotypes and has been reported in studies of tumor biology, supporting its relevance to pathways that govern proliferation, differentiation, and cellular identity. These features make Zfhx4 a useful target for interrogating transcriptional circuitry and disease-associated gene regulatory changes in mouse model systems.

ZFHX4 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Zfhx4 expression without altering the underlying DNA sequence.

ZFHX4 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Zfhx4 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 Zfhx4 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous ZFHX4 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Zfhx4 locus and enabling the study of ZFHX4-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of ZFHX4 pathway restoration in tumor cells with silenced or reduced Zfhx4 expression.

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