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.
Polycystin-2 CRISPR/Cas9 KO Plasmid (h): sc-400669
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Polycystin-2 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 Polycystin-2 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: Polycystin-2 Antibody (YCE2): sc-47734
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Polycystin-2 CRISPR/Cas9 KO Plasmid (h) | sc-400669 | 20 µg | $397.00 |
Overview
PKD2 encodes polycystin-2, a Ca2+-permeable nonselective cation channel of the TRP family that localizes to primary cilia, endoplasmic reticulum, and plasma membrane to coordinate mechanosensory and intracellular calcium signaling. Polycystin-2 forms functional complexes with polycystin-1 and contributes to cilia-dependent pathways that regulate epithelial polarity, tubular morphogenesis, and cell-cycle control through Ca2+-linked effectors. Disruption of PKD2 perturbs calcium homeostasis and downstream signaling programs that influence proliferation, differentiation, and fluid transport in renal and other epithelia. Genetic alteration of PKD2 is strongly associated with autosomal dominant polycystic kidney disease, making it a key node for interrogating ciliary signaling and cystogenesis-related mechanisms in human cell models.
Polycystin-2 CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the PKD2 gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the PKD2 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 PKD2 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 Polycystin-2 protein expression.
This CRISPR knockout system enables efficient generation of PKD2-deficient cell models for investigation of Polycystin-2 signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting PKD2 exon(s) critical for Polycystin-2 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 PKD2 genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by Polycystin-2 CRISPR/Cas9 KO Plasmid (h) and Polycystin-2 CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the PKD2 locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by Polycystin-2 HDR Plasmid (h) and Polycystin-2 HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by PKD2 homology arms to support homology-directed repair at defined PKD2 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.