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.
nephrin CRISPR/Cas9 KO Plasmid (h): sc-400368
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- nephrin 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 nephrin 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: nephrin Antibody (G-8): sc-376522
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
nephrin CRISPR/Cas9 KO Plasmid (h) | sc-400368 | 20 µg | $397.00 |
Overview
NPHS1 encodes nephrin, a transmembrane immunoglobulin superfamily protein that is essential for the architecture and signaling functions of the podocyte slit diaphragm in the renal glomerulus. Nephrin participates in cell–cell junction organization and actin cytoskeleton regulation through interactions with adaptor proteins and downstream phosphorylation-dependent pathways, helping maintain filtration barrier integrity. Disruption of NPHS1 compromises slit diaphragm structure, alters podocyte adhesion and survival signaling, and is linked to proteinuric kidney phenotypes including congenital nephrotic syndrome. NPHS1 is therefore a key target for investigating podocyte biology, filtration barrier dynamics, and junctional signaling mechanisms.
nephrin CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the NPHS1 gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the NPHS1 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 NPHS1 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 nephrin protein expression.
This CRISPR knockout system enables efficient generation of NPHS1-deficient cell models for investigation of nephrin signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting NPHS1 exon(s) critical for nephrin 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 NPHS1 genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by nephrin CRISPR/Cas9 KO Plasmid (h) and nephrin CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the NPHS1 locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by nephrin HDR Plasmid (h) and nephrin HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by NPHS1 homology arms to support homology-directed repair at defined NPHS1 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.