PLU-1 CRISPR Activation Plasmid (h): sc-403771-ACT

KDM5B encodes PLU-1 (JARID1B), a Jumonji C (JmjC) domain–containing histone lysine demethylase that preferentially removes H3K4me3/2 marks, thereby reshaping promoter-associated chromatin and transcriptional programs. Through interactions with lineage-specific transcription factors and chromatin remodeling complexes, PLU-1 contributes to regulation of cell cycle progression, differentiation states, and replication-linked chromatin maintenance. Altered KDM5B activity has been linked to epigenetic plasticity and transcriptional rewiring in multiple tumor contexts, making it a widely used target for studying chromatin-dependent control of proliferation and cell identity. In human cell models, perturbing PLU-1 helps dissect pathways connecting histone methylation dynamics to gene expression outputs and phenotypic transitions.

PLU-1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous KDM5B expression without altering the underlying DNA sequence.

PLU-1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the KDM5B 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 KDM5B transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous PLU-1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native KDM5B locus and enabling the study of PLU-1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of PLU-1 pathway restoration in tumor cells with silenced or reduced KDM5B expression.

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