Not CRISPR data: the image illustrates antibody quality for visualizing CRISPR experiment results.
Not CRISPR data: the image illustrates antibody quality for visualizing CRISPR experiment results.
DARPP-32 CRISPR/Cas9 KO Plasmid (m): sc-422380
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- DARPP-32 CRISPR/Cas9 Knockout (KO) Plasmid (m) is a pool of plasmids, each encoding Cas9 nuclease and a target-specific 20 nt guide RNA (gRNA) designed for maximum knockout efficiency using sequences derived from the GeCKO v2 library
- gRNA sequences direct Cas9 to induce site-specific double-strand breaks (DSBs) in the DARPP-32 genomic locus, resulting in gene knockout through non-homologous end joining (NHEJ)
- The puromycin resistance and RFP genes are flanked by LoxP sites, enabling removal of selection markers via Cre recombinase (Cre Vector: sc-418923) after establishing stable knockout cell lines
- 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/Cas9 KO Plasmid (m) | sc-422380 | 20 µg | $397.00 |
Overview
Ppp1r1b encodes DARPP-32 (dopamine- and cAMP-regulated phosphoprotein, 32 kDa), a pivotal integrator of dopaminergic and glutamatergic signaling in striatal medium spiny neurons. Through phosphorylation-dependent switching, DARPP-32 modulates protein phosphatase-1 activity and links cAMP/PKA signaling with downstream regulation of synaptic plasticity, ion channel function, and neurotransmitter receptor signaling. This node intersects pathways governing reward learning, motor control, and transcriptional responses to neuromodulators, making Ppp1r1b a common readout in studies of basal ganglia circuitry. Altered DARPP-32 signaling has been associated with neuropsychiatric and neurodegenerative disease mechanisms, including dysregulated dopamine signaling, maladaptive plasticity, and stress-related behavioral phenotypes in mouse models.
DARPP-32 CRISPR/Cas9 KO Plasmid (m) is a pool of plasmids designed for targeted disruption of the Ppp1r1b gene in mouse cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the Ppp1r1b together with the Streptococcus pyogenes Cas9 nuclease. The plasmids also encode GFP, allowing fluorescent identification and enrichment of successfully transfected cells by fluorescence microscopy or flow cytometry.
The multi-guide design increases the likelihood of generating insertions or deletions (indels) that disrupt the Ppp1r1b open reading frame following Cas9-mediated double-strand break formation. DNA breaks introduced by the CRISPR/Cas9 system are repaired through endogenous non-homologous end joining (NHEJ) pathways, frequently resulting in frameshift mutations that abolish DARPP-32 protein expression.
This CRISPR knockout system enables efficient generation of Ppp1r1b-deficient cell models for investigation of DARPP-32 signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting Ppp1r1b exon(s) critical for DARPP-32 function
- Co-expression of SpCas9 and sgRNA from a single plasmid for simplified delivery
- GFP reporter for identification of transfected cells
- Pool of plasmids targeting multiple Ppp1r1b genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by DARPP-32 CRISPR/Cas9 KO Plasmid (m) and DARPP-32 CRISPR/Cas9 KO Plasmid (m2) target distinct sites within the Ppp1r1b locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by DARPP-32 HDR Plasmid (m) and DARPP-32 HDR Plasmid (m2) contain a puromycin resistance cassette and an RFP reporter flanked by Ppp1r1b homology arms to support homology-directed repair at defined Ppp1r1b target sites corresponding to the CRISPR/Cas9 KO designs. HDR donor availability may vary. See Related Products for availability.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.