JMJD3 CRISPR Activation Plasmid (h): sc-401883-ACT

Human KDM6B encodes the histone lysine demethylase JMJD3, an Fe(II)/2-oxoglutarate–dependent enzyme that removes repressive H3K27me3 marks to promote transcriptional programs linked to cell fate transitions. JMJD3 integrates with chromatin remodeling and Polycomb-regulated gene networks to control lineage commitment, inflammatory gene expression, and stimulus-responsive transcription. Through these epigenetic mechanisms, KDM6B contributes to regulation of differentiation, senescence, and immune signaling pathways, making it relevant to studies of developmental disorders, cancer-associated transcriptional reprogramming, and chronic inflammatory states. Perturbation of JMJD3 activity or expression can shift enhancer and promoter landscapes, influencing downstream pathways such as NF-κB–responsive transcription and context-dependent oncogenic or tumor-suppressive programs.

JMJD3 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous KDM6B expression without altering the underlying DNA sequence.

JMJD3 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the KDM6B 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 KDM6B transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous JMJD3 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native KDM6B locus and enabling the study of JMJD3-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of JMJD3 pathway restoration in tumor cells with silenced or reduced KDM6B expression.

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