TET3 CRISPR Activation Plasmid (h): sc-414997-ACT

Human TET3 encodes a Fe(II)/2-oxoglutarate–dependent dioxygenase that catalyzes iterative oxidation of 5-methylcytosine to 5-hydroxymethylcytosine and downstream oxidized bases, supporting active and passive DNA demethylation. Through this epigenetic editing activity, TET3 regulates transcriptional programs involved in chromatin remodeling, lineage specification, and cellular stress responses. TET3 function intersects with DNA repair–coupled demethylation processes and broader chromatin-state maintenance pathways that shape cell identity. Dysregulated TET3 activity and altered 5hmC patterns have been associated with developmental and neurological phenotypes and are frequently studied in the context of epigenetic instability in cancer biology.

TET3 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous TET3 expression without altering the underlying DNA sequence.

TET3 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the TET3 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 TET3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous TET3 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native TET3 locus and enabling the study of TET3-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of TET3 pathway restoration in tumor cells with silenced or reduced TET3 expression.

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