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.
NEDD4-L CRISPR/Cas9 KO Plasmid (m): sc-429949
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- NEDD4-L 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 NEDD4-L 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: NEDD4-L Antibody (C-8): sc-514954
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
NEDD4-L CRISPR/Cas9 KO Plasmid (m) | sc-429949 | 20 µg | $397.00 |
Overview
Nedd4l encodes NEDD4-L, a HECT-domain E3 ubiquitin ligase that regulates protein turnover by catalyzing ubiquitination of membrane and signaling proteins, including ion channels and transporters such as ENaC. Through modulation of ubiquitin-dependent endocytosis and lysosomal/proteasomal degradation, NEDD4-L influences epithelial ion homeostasis and signal transduction pathways implicated in cellular stress responses and tissue remodeling. In mouse systems, altered Nedd4l activity has been linked to dysregulated sodium handling and blood pressure control, and has been investigated in contexts of renal physiology, cardiovascular phenotypes, and inflammatory signaling. NEDD4-L also interfaces with pathways controlling cell polarity and differentiation, making it relevant for studies of epithelial barrier function and developmental processes.
NEDD4-L CRISPR/Cas9 KO Plasmid (m) is a pool of plasmids designed for targeted disruption of the Nedd4l gene in mouse cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the Nedd4l 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 Nedd4l 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 NEDD4-L protein expression.
This CRISPR knockout system enables efficient generation of Nedd4l-deficient cell models for investigation of NEDD4-L signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting Nedd4l exon(s) critical for NEDD4-L 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 Nedd4l genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by NEDD4-L CRISPR/Cas9 KO Plasmid (m) and NEDD4-L CRISPR/Cas9 KO Plasmid (m2) target distinct sites within the Nedd4l locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by NEDD4-L HDR Plasmid (m) and NEDD4-L HDR Plasmid (m2) contain a puromycin resistance cassette and an RFP reporter flanked by Nedd4l homology arms to support homology-directed repair at defined Nedd4l 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.