TET1 CRISPR Activation Plasmid (m): sc-424616-ACT

Mouse Tet1 encodes the dioxygenase TET1, a key epigenetic regulator that catalyzes oxidation of 5-methylcytosine to 5-hydroxymethylcytosine and related derivatives, promoting DNA demethylation and shaping chromatin accessibility. TET1 activity influences transcriptional programs governing embryonic development, lineage commitment, neuronal gene regulation, and maintenance of cellular identity through coordination with methylation and chromatin remodeling pathways. By modulating CpG methylation landscapes, TET1 impacts genome stability and enhancer/promoter function, linking it to dysregulated differentiation and aberrant gene expression states observed across diverse disease-relevant models. In mouse systems, Tet1 perturbation is commonly studied in the context of neurodevelopment, stem cell biology, and epigenome dynamics that affect cellular stress responses and inflammatory signaling.

TET1 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Tet1 expression without altering the underlying DNA sequence.

TET1 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Tet1 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 Tet1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous TET1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Tet1 locus and enabling the study of TET1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of TET1 pathway restoration in tumor cells with silenced or reduced Tet1 expression.

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