Dnmt3b CRISPR Activation Plasmid (m): sc-420036-ACT
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Dnmt3b CRISPR Activation Plasmid (m) is a synergistic activation mediator (SAM) transcription activation system designed to specifically upregulate gene expression
- Dnmt3b CRISPR Activation Plasmid (m) consists of three plasmids at a 1:1:1 mass ratio: a plasmid encoding the deactivated Cas9 (dCas9) nuclease (D10A and N863A) fused to the transactivation domain VP64, and a blasticidin resistance gene; a plasmid encoding the MS2-p65-HSF1 fusion protein, and a hygromycin resistance gene; a plasmid encoding a target-specific 20 nt guide RNA fused to two MS2 RNA aptamers, and a puromycin resistance gene
- The resulting SAM complex binds to a site-specific region approximately 200-250 nt upstream of the transcriptional start site and provides robust recruitment of transcription factors for highly efficient gene activation
- gRNAs encoded by Dnmt3b CRISPR Activation Plasmid (m) and Dnmt3b CRISPR Activation Plasmid (m2) target distinct regulatory regions upstream of the Dnmt3b transcriptional start site. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: Dnmt3b Antibody (G-9): sc-376043
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Dnmt3b CRISPR Activation Plasmid (m) | sc-420036-ACT | 20 µg | $397.00 | |||
Dnmt3b CRISPR Activation Plasmid (m2) | sc-420036-ACT-2 | 20 µg | $397.00 |
Mouse Dnmt3b encodes a de novo DNA methyltransferase that deposits 5-methylcytosine at CpG dinucleotides to establish and maintain epigenetic silencing during embryonic development and lineage commitment. DNMT3B cooperates with DNMT3A and UHRF1/DNMT1-mediated maintenance methylation to shape chromatin state, transcriptional programs, and genomic stability, including repression of transposable elements and regulation of imprinting. This enzyme integrates with chromatin remodeling and histone modification pathways, influencing promoter and enhancer methylation dynamics across differentiation and reprogramming. Altered DNMT3B activity is linked to aberrant methylation patterns observed in developmental disorders and cancer-associated epigenetic dysregulation, making it a central node for studying epigenome-dependent gene regulation.
Dnmt3b CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Dnmt3b expression without altering the underlying DNA sequence.
Dnmt3b CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Dnmt3b 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 Dnmt3b transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Dnmt3b expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Dnmt3b locus and enabling the study of Dnmt3b-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Dnmt3b pathway restoration in tumor cells with silenced or reduced Dnmt3b expression.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.