Dnmt1 CRISPR Activation Plasmid (m): sc-420033-ACT

Mouse Dnmt1 encodes DNA methyltransferase 1, the primary maintenance methyltransferase that copies CpG methylation patterns during DNA replication to preserve epigenetic cell identity. DNMT1 functions within chromatin regulatory networks alongside UHRF1 and methyl-CpG binding proteins, influencing transcriptional repression, genomic imprinting, X-chromosome inactivation, and transposon silencing. Through coupling DNA methylation with histone modifications and replication-coupled chromatin assembly, DNMT1 helps maintain genome stability and coordinated gene expression programs. Dysregulated DNMT1 activity or expression is linked to aberrant methylation landscapes implicated in oncogenic transformation, developmental defects, and neurobiological phenotypes, making it a central node for epigenetics and disease-model research.

Dnmt1 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Dnmt1 expression without altering the underlying DNA sequence.

Dnmt1 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Dnmt1 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 Dnmt1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Dnmt1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Dnmt1 locus and enabling the study of Dnmt1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Dnmt1 pathway restoration in tumor cells with silenced or reduced Dnmt1 expression.

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