FBL10 CRISPR Activation Plasmid (h): sc-403232-ACT

KDM2B (also known as FBL10) encodes a JmjC domain–containing histone demethylase that functions in epigenetic regulation of transcription through chromatin remodeling. The protein is implicated in Polycomb-associated gene repression and modulation of cell-cycle and differentiation programs via control of promoter-associated histone methylation states. Through these activities, KDM2B/FBL10 influences stemness maintenance, lineage commitment, and cellular senescence, linking its dysregulation to altered proliferative signaling and aberrant transcriptional networks. Perturbations in KDM2B expression or function have been reported across cancer-relevant contexts and developmental phenotypes, making it a useful node for studying chromatin-dependent control of gene expression.

FBL10 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous KDM2B expression without altering the underlying DNA sequence.

FBL10 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the KDM2B 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 KDM2B transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous FBL10 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native KDM2B locus and enabling the study of FBL10-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of FBL10 pathway restoration in tumor cells with silenced or reduced KDM2B expression.

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