SmcX CRISPR Activation Plasmid (m): sc-423030-ACT

Kdm5c (SmcX) encodes an X-linked Jumonji C (JmjC) domain–containing histone demethylase that removes H3K4me2/3 marks, shaping chromatin accessibility and transcriptional programs during development. In mouse cells, SmcX modulates epigenetic control of gene expression linked to neuronal differentiation, synaptic function, and activity-dependent transcription, interfacing with pathways that regulate cell fate decisions and genome-wide promoter/enhancer states. Disruption or dysregulation of KDM5C is associated with neurodevelopmental phenotypes and altered cognitive function, consistent with its role in maintaining appropriate transcriptional homeostasis. SmcX is therefore a useful target for studying chromatin-driven regulation, neuronal gene networks, and epigenetic contributions to disease-relevant cellular states.

SmcX CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Kdm5c expression without altering the underlying DNA sequence.

SmcX CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Kdm5c 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 Kdm5c transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous SmcX expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Kdm5c locus and enabling the study of SmcX-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of SmcX pathway restoration in tumor cells with silenced or reduced Kdm5c expression.

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