MZF-1 CRISPR Activation Plasmid (h): sc-403142-ACT

Myeloid zinc finger 1 (MZF1/MZF-1) is a KRAB-domain C2H2 zinc finger transcription factor with prominent roles in hematopoietic lineage commitment and myeloid differentiation. It regulates gene expression programs that influence cell-cycle progression, apoptosis, and cellular migration, integrating with broader transcriptional networks controlling proliferation and stress responses. Dysregulated MZF1 activity has been associated with altered differentiation states and aberrant transcriptional reprogramming observed in hematologic disorders and multiple solid tumor contexts. As a DNA-binding regulator, MZF-1 is frequently studied for its impact on oncogenic signaling, metastatic potential, and lineage-specific transcriptional circuitry.

MZF-1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous MZF1 expression without altering the underlying DNA sequence.

MZF-1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the MZF1 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 MZF1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous MZF-1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native MZF1 locus and enabling the study of MZF-1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of MZF-1 pathway restoration in tumor cells with silenced or reduced MZF1 expression.

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