Podocin CRISPR/Cas9 KO Plasmid (h): sc-400930
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Podocin 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 Podocin 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 | |
Podocin CRISPR/Cas9 KO Plasmid (h) | sc-400930 | 20 µg | $397.00 |
Overview
NPHS2 encodes podocin, an essential slit diaphragm protein in human podocytes that scaffolds signaling complexes at the glomerular filtration barrier. Podocin interacts with nephrin and CD2AP and helps organize lipid raft microdomains, supporting cytoskeletal dynamics, mechanosensing, and maintenance of foot process architecture. Disruption of NPHS2 perturbs slit diaphragm integrity and alters podocyte signaling pathways linked to actin remodeling and cell survival. Genetic and functional defects in podocin are strongly associated with proteinuric kidney disease, including steroid-resistant nephrotic syndrome and focal segmental glomerulosclerosis, making NPHS2 a key target for modeling glomerular barrier dysfunction.
Podocin CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the NPHS2 gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the NPHS2 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 NPHS2 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 Podocin protein expression.
This CRISPR knockout system enables efficient generation of NPHS2-deficient cell models for investigation of Podocin signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting NPHS2 exon(s) critical for Podocin 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 NPHS2 genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by Podocin CRISPR/Cas9 KO Plasmid (h) and Podocin CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the NPHS2 locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by Podocin HDR Plasmid (h) and Podocin HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by NPHS2 homology arms to support homology-directed repair at defined NPHS2 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.