NIF-1 CRISPR Activation Plasmid (h): sc-412963-ACT

Human ZNF335 encodes NIF-1, a nuclear zinc finger protein that functions as a chromatin-associated regulator of transcription and genome-wide gene expression programs. NIF-1 has been linked to the control of neural progenitor proliferation and differentiation through coordination of epigenetic and transcriptional networks that shape developmental gene sets. By influencing chromatin state and promoter activity, ZNF335/NIF-1 intersects with pathways governing cell-cycle progression, lineage specification, and maintenance of cellular identity. Dysregulated ZNF335 expression or function has been associated with neurodevelopmental abnormalities and provides a mechanistic entry point for studying transcriptional control in disease-relevant models.

NIF-1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ZNF335 expression without altering the underlying DNA sequence.

NIF-1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ZNF335 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 ZNF335 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous NIF-1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ZNF335 locus and enabling the study of NIF-1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of NIF-1 pathway restoration in tumor cells with silenced or reduced ZNF335 expression.

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