Jade-1 CRISPR Activation Plasmid (h): sc-410620-ACT

JADE1 encodes Jade-1, a nuclear PHD zinc finger protein that functions as an epigenetic regulator by recognizing histone marks and partnering with histone acetyltransferase complexes to modulate chromatin accessibility and transcription. Through these interactions, Jade-1 influences programs linked to cell-cycle control, DNA damage responses, and differentiation-associated gene expression. JADE1 has also been studied in relation to ubiquitin-dependent regulation of signaling proteins and transcription factors, connecting chromatin remodeling to proteostasis. Altered JADE1 expression or dysregulated Jade-1–associated chromatin pathways has been implicated in contexts of aberrant proliferation and tumor-associated transcriptional reprogramming, supporting its utility as a mechanistic node for pathway-focused research.

Jade-1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous JADE1 expression without altering the underlying DNA sequence.

Jade-1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the JADE1 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 JADE1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Jade-1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native JADE1 locus and enabling the study of Jade-1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Jade-1 pathway restoration in tumor cells with silenced or reduced JADE1 expression.

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