ZFHX4 CRISPR Activation Plasmid (h): sc-408957-ACT

ZFHX4 (zinc finger homeobox 4) encodes a large nuclear transcription factor with multiple zinc finger and homeobox DNA-binding domains that modulates gene expression programs controlling cell identity and differentiation. It participates in transcriptional regulation networks linked to neurodevelopmental and epithelial lineage processes, influencing chromatin state and downstream signaling outputs. Dysregulated ZFHX4 expression or mutation has been associated with altered differentiation trajectories and tumor biology in several contexts, supporting its use as a molecular node for studying oncogenic transcriptional remodeling. Functional interrogation of ZFHX4 helps clarify how transcription factors integrate developmental cues with cell cycle control and stress-adaptive pathways.

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

ZFHX4 CRISPR Activation Plasmid (h) 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.