RBP2 CRISPR Activation Plasmid (h): sc-403940-ACT

KDM5A encodes RBP2, a JmjC domain–containing histone lysine demethylase that preferentially removes H3K4me3/me2 marks to regulate promoter activity and transcriptional output. By shaping chromatin accessibility and RNA polymerase II–dependent gene expression programs, RBP2 influences cell-cycle progression, lineage specification, and responses to developmental and stress cues. Its activity integrates with epigenetic control networks involving enhancer–promoter regulation and transcription factor occupancy. Dysregulated KDM5A/RBP2 function has been associated with aberrant transcriptional states observed across multiple disease-relevant contexts, including cancer biology and neurodevelopmental phenotypes, making it a useful node for mechanistic studies of chromatin-driven gene regulation.

RBP2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous KDM5A expression without altering the underlying DNA sequence.

RBP2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the KDM5A 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 KDM5A transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous RBP2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native KDM5A locus and enabling the study of RBP2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of RBP2 pathway restoration in tumor cells with silenced or reduced KDM5A expression.

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