DARPP-32 CRISPR Activation Plasmid (h): sc-400430-ACT
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- DARPP-32 CRISPR Activation Plasmid (h) is a synergistic activation mediator (SAM) transcription activation system designed to specifically upregulate gene expression
- DARPP-32 CRISPR Activation Plasmid (h) 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 DARPP-32 CRISPR Activation Plasmid (h) and DARPP-32 CRISPR Activation Plasmid (h2) target distinct regulatory regions upstream of the PPP1R1B 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: DARPP-32 Antibody (H-3): sc-271111
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
DARPP-32 CRISPR Activation Plasmid (h) | sc-400430-ACT | 20 µg | $397.00 | |||
DARPP-32 CRISPR Activation Plasmid (h2) | sc-400430-ACT-2 | 20 µg | $397.00 |
PPP1R1B encodes DARPP-32 (dopamine- and cAMP-regulated phosphoprotein of 32 kDa), a pivotal integrator of dopaminergic and glutamatergic signaling in neurons. When phosphorylated by PKA, DARPP-32 inhibits protein phosphatase 1, thereby modulating downstream phosphorylation states that shape synaptic plasticity, transcriptional responses, and neuronal excitability. This regulation links PPP1R1B to cAMP/PKA signaling, calcium-dependent pathways, and receptor-driven phosphorylation networks in striatal medium spiny neurons. Altered DARPP-32 expression or phosphorylation is frequently studied in the context of neuropsychiatric and neurodegenerative disease mechanisms, as well as signal transduction changes in tumor biology.
DARPP-32 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PPP1R1B expression without altering the underlying DNA sequence.
DARPP-32 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PPP1R1B 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 PPP1R1B transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous DARPP-32 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PPP1R1B locus and enabling the study of DARPP-32-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of DARPP-32 pathway restoration in tumor cells with silenced or reduced PPP1R1B expression.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.