SMYD3 CRISPR Activation Plasmid (h): sc-402128-ACT

SMYD3 (SET and MYND domain-containing protein 3) is a lysine methyltransferase that catalyzes histone H3 methylation, most prominently at H3K4, to promote transcriptional activation of target gene programs. Through chromatin remodeling and regulation of promoter-proximal transcription, SMYD3 influences core processes including cell-cycle progression, DNA damage response, and differentiation. SMYD3 also methylates select non-histone substrates, linking epigenetic control to signaling and cytoskeletal dynamics. Dysregulated SMYD3 expression and activity have been associated with oncogenic transcriptional networks and altered proliferative phenotypes, making it a useful target for mechanistic studies of epigenetic regulation in human cells.

SMYD3 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SMYD3 expression without altering the underlying DNA sequence.

SMYD3 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SMYD3 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 SMYD3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous SMYD3 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SMYD3 locus and enabling the study of SMYD3-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of SMYD3 pathway restoration in tumor cells with silenced or reduced SMYD3 expression.

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