Pr-Set7 CRISPR Activation Plasmid (h): sc-415759-ACT

KMT5A encodes Pr-Set7 (SETD8), a SET domain–containing lysine methyltransferase that catalyzes monomethylation of histone H4 at Lys20 (H4K20me1), a chromatin mark linked to replication-coupled nucleosome maturation and genome integrity. Pr-Set7 activity coordinates cell-cycle progression, DNA replication licensing, and DNA damage responses through regulation of chromatin compaction and recruitment of repair factors. Dysregulated KMT5A/Pr-Set7 expression or activity has been associated with altered epigenetic states, replication stress, and proliferative phenotypes reported across multiple cancer contexts. As a chromatin enzyme at the intersection of epigenetic control and checkpoint signaling, KMT5A is widely studied in transcriptional regulation, DNA repair pathway crosstalk, and chromatin dynamics.

Pr-Set7 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous KMT5A expression without altering the underlying DNA sequence.

Pr-Set7 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the KMT5A 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 KMT5A transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Pr-Set7 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native KMT5A locus and enabling the study of Pr-Set7-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Pr-Set7 pathway restoration in tumor cells with silenced or reduced KMT5A expression.

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