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.
NNT CRISPR/Cas9 KO Plasmid (m): sc-421922
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- NNT 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 NNT 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: NNT Antibody (B-3): sc-390236
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
NNT CRISPR/Cas9 KO Plasmid (m) | sc-421922 | 20 µg | $397.00 |
Overview
Mouse Nnt encodes nicotinamide nucleotide transhydrogenase (NNT), a mitochondrial inner-membrane enzyme that couples proton motive force to hydride transfer between NAD(H) and NADP(H). By generating mitochondrial NADPH, NNT supports glutathione and thioredoxin-dependent antioxidant systems and helps maintain redox homeostasis during oxidative phosphorylation. NNT activity influences mitochondrial metabolism, reactive oxygen species handling, and susceptibility to oxidative stress–driven dysfunction. Altered Nnt/NNT function has been linked to metabolic phenotypes and impaired stress responses, making it relevant for studies of mitochondrial bioenergetics and redox-regulated signaling.
NNT CRISPR/Cas9 KO Plasmid (m) is a pool of plasmids designed for targeted disruption of the Nnt gene in mouse cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the Nnt 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 Nnt 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 NNT protein expression.
This CRISPR knockout system enables efficient generation of Nnt-deficient cell models for investigation of NNT signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting Nnt exon(s) critical for NNT 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 Nnt genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by NNT CRISPR/Cas9 KO Plasmid (m) and NNT CRISPR/Cas9 KO Plasmid (m2) target distinct sites within the Nnt locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by NNT HDR Plasmid (m) and NNT HDR Plasmid (m2) contain a puromycin resistance cassette and an RFP reporter flanked by Nnt homology arms to support homology-directed repair at defined Nnt 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.