TET1 CRISPR Activation Plasmid (h): sc-400845-ACT

Human TET1 (ten-eleven translocation methylcytosine dioxygenase 1) is a Fe(II)/2-oxoglutarate–dependent dioxygenase that catalyzes oxidation of 5-methylcytosine to 5-hydroxymethylcytosine and further oxidized derivatives, initiating active DNA demethylation. Through this activity, TET1 shapes epigenetic regulation of transcription, chromatin state, and developmental gene programs, influencing processes such as lineage specification, genome stability, and DNA damage responses. TET1-dependent remodeling of DNA methylation patterns interfaces with transcriptional networks and chromatin modifiers to control enhancer and promoter activity. Dysregulated TET1 expression or altered 5hmC landscapes have been associated with aberrant transcriptional states in cancer and with epigenetic alterations relevant to neurodevelopment and immune regulation, making it a key target for mechanistic studies of epigenome-driven phenotypes.

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

TET1 CRISPR Activation Plasmid (h) 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.