ZFP100 CRISPR Activation Plasmid (h): sc-411821-ACT

Human ZNF473 encodes the zinc finger protein ZFP100, a putative DNA-binding transcriptional regulator characterized by C2H2-type zinc finger domains that are commonly linked to sequence-specific control of gene expression. By modulating promoter and enhancer activity, ZFP100 is expected to influence chromatin-dependent transcriptional programs that shape cell identity, stress responses, and differentiation-related pathways. Altered regulation of zinc finger transcription factors is frequently associated with dysregulated epigenetic states and aberrant signaling networks, making ZNF473 a useful locus for investigating mechanisms of transcriptional control in disease-relevant contexts. Characterizing ZFP100-dependent gene networks can support studies of pathway remodeling, regulatory element function, and transcriptomic changes in human cells.

ZFP100 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ZNF473 expression without altering the underlying DNA sequence.

ZFP100 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ZNF473 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 ZNF473 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous ZFP100 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ZNF473 locus and enabling the study of ZFP100-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of ZFP100 pathway restoration in tumor cells with silenced or reduced ZNF473 expression.

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