ZNF649 CRISPR Activation Plasmid (h): sc-418512-ACT

ZNF649 encodes a C2H2-type zinc finger transcription factor implicated in sequence-specific DNA binding and regulation of gene expression programs that influence cellular identity and chromatin-associated processes. As a nuclear regulatory protein, ZNF649 is often considered within broader KRAB zinc finger–linked networks that modulate transcriptional repression/activation dynamics, potentially affecting cell cycle progression, differentiation, and stress-response pathways. Altered regulation of zinc finger transcription factors has been associated with dysregulated transcriptional circuitry in cancer and other diseases characterized by aberrant epigenetic control, making ZNF649 relevant for mechanistic studies of transcriptional homeostasis. Investigating ZNF649 activity can help clarify how promoter- and enhancer-associated binding events shape downstream signaling and gene regulatory networks in human cells.

ZNF649 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ZNF649 expression without altering the underlying DNA sequence.

ZNF649 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ZNF649 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 ZNF649 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous ZNF649 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ZNF649 locus and enabling the study of ZNF649-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of ZNF649 pathway restoration in tumor cells with silenced or reduced ZNF649 expression.

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