pki α CRISPR/Cas9 KO Plasmid (h): sc-416391
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- pki α CRISPR/Cas9 Knockout (KO) Plasmid (h) 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 pki α 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
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
pki α CRISPR/Cas9 KO Plasmid (h) | sc-416391 | 20 µg | $397.00 |
Overview
PKIA encodes protein kinase inhibitor alpha (PKIα), a heat-stable inhibitor that binds the catalytic subunit of cAMP-dependent protein kinase A (PKA) and suppresses kinase activity. By restraining cAMP/PKA signaling, PKIα influences phosphorylation programs controlling transcriptional regulation, cell-cycle progression, metabolism, and stress responses, including modulation of CREB-dependent gene expression. PKIα also contributes to spatial control of PKA signaling through interactions that affect kinase localization and signaling amplitude. Dysregulated cAMP/PKA pathway activity and altered PKIA expression have been associated with contexts relevant to tumor biology, endocrine signaling, and neuronal function, supporting its use in pathway-centric mechanistic studies.
pki α CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the PKIA gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the PKIA 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 PKIA 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 pki α protein expression.
This CRISPR knockout system enables efficient generation of PKIA-deficient cell models for investigation of pki α signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting PKIA exon(s) critical for pki α 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 PKIA genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by pki α CRISPR/Cas9 KO Plasmid (h) and pki α CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the PKIA locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by pki α HDR Plasmid (h) and pki α HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by PKIA homology arms to support homology-directed repair at defined PKIA 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.