Notum CRISPR/Cas9 KO Plasmid (h): sc-409374
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Notum 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 Notum 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 | |
Notum CRISPR/Cas9 KO Plasmid (h) | sc-409374 | 20 µg | $397.00 |
Overview
NOTUM encodes Notum, a secreted carboxylesterase that negatively regulates Wnt signaling by deacylating the palmitoleate modification on Wnt ligands, thereby limiting ligand–receptor engagement and downstream β-catenin–dependent transcription. By shaping Wnt gradient formation and signal duration, Notum contributes to developmental patterning, tissue homeostasis, and stem cell niche regulation. Altered NOTUM activity has been linked to dysregulated Wnt pathway output observed across multiple disease contexts, including cancer biology and disorders involving aberrant epithelial renewal and differentiation. As a soluble Wnt antagonist, Notum is also relevant to studies of extracellular matrix–proximal signaling and paracrine control of pathway activation.
Notum CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the NOTUM gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the NOTUM 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 NOTUM 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 Notum protein expression.
This CRISPR knockout system enables efficient generation of NOTUM-deficient cell models for investigation of Notum signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting NOTUM exon(s) critical for Notum 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 NOTUM genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by Notum CRISPR/Cas9 KO Plasmid (h) and Notum CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the NOTUM locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by Notum HDR Plasmid (h) and Notum HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by NOTUM homology arms to support homology-directed repair at defined NOTUM 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.