NCOAT CRISPR/Cas9 KO Plasmid (m): sc-429251
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- NCOAT 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 NCOAT 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 | |
NCOAT CRISPR/Cas9 KO Plasmid (m) | sc-429251 | 20 µg | $397.00 |
Overview
Mgea5 encodes NCOAT (also known as O‑GlcNAcase), a key enzyme that removes O‑linked β‑N‑acetylglucosamine (O‑GlcNAc) from serine and threonine residues on nuclear and cytoplasmic proteins. Together with O‑GlcNAc transferase, NCOAT regulates dynamic O‑GlcNAc cycling that integrates nutrient sensing with phosphorylation-dependent signaling, influencing transcription, cell-cycle progression, proteostasis, and stress responses. NCOAT activity impacts pathways such as insulin/AKT signaling, chromatin regulation, and protein quality control by modulating O‑GlcNAc levels on transcription factors, kinases, and cytoskeletal proteins. Dysregulated O‑GlcNAc homeostasis has been linked to metabolic and neurodegeneration-associated phenotypes, making Mgea5 a useful target in mouse models of signaling and epigenetic regulation.
NCOAT CRISPR/Cas9 KO Plasmid (m) is a pool of plasmids designed for targeted disruption of the Mgea5 gene in mouse cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the Mgea5 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 Mgea5 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 NCOAT protein expression.
This CRISPR knockout system enables efficient generation of Mgea5-deficient cell models for investigation of NCOAT signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting Mgea5 exon(s) critical for NCOAT 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 Mgea5 genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by NCOAT CRISPR/Cas9 KO Plasmid (m) and NCOAT CRISPR/Cas9 KO Plasmid (m2) target distinct sites within the Mgea5 locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by NCOAT HDR Plasmid (m) and NCOAT HDR Plasmid (m2) contain a puromycin resistance cassette and an RFP reporter flanked by Mgea5 homology arms to support homology-directed repair at defined Mgea5 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.