JHDM1D CRISPR Activation Plasmid (h): sc-405142-ACT

KDM7A (also known as JHDM1D) encodes a Jumonji C domain–containing histone demethylase that modulates chromatin accessibility by removing repressive methyl marks on lysine residues of histone H3, thereby shaping transcriptional programs. Through epigenetic regulation, JHDM1D influences processes such as cell fate specification, lineage-associated gene expression, and coordination of developmental and stress-responsive transcriptional networks. Dysregulated histone methylation dynamics involving KDM7A have been implicated in aberrant transcriptional states relevant to oncogenic transformation and neurodevelopmental phenotypes, making it a useful node for mechanistic studies of chromatin-driven gene control. Its activity intersects with broader epigenetic pathways that couple histone modification status to RNA polymerase II–dependent transcription and cellular identity maintenance.

JHDM1D CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous KDM7A expression without altering the underlying DNA sequence.

JHDM1D CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the KDM7A 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 KDM7A transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous JHDM1D expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native KDM7A locus and enabling the study of JHDM1D-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of JHDM1D pathway restoration in tumor cells with silenced or reduced KDM7A expression.

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