ME3 CRISPR/Cas9 KO Plasmid (h): sc-417146
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- ME3 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 ME3 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 | |
ME3 CRISPR/Cas9 KO Plasmid (h) | sc-417146 | 20 µg | $397.00 |
Overview
Human ME3 encodes the mitochondrial NADP-dependent malic enzyme that catalyzes oxidative decarboxylation of malate to pyruvate with concomitant NADPH production. By linking the tricarboxylic acid cycle to mitochondrial redox control, ME3 supports biosynthetic reducing power, antioxidant defense, and metabolic flexibility under changing nutrient conditions. ME3 activity influences pathways involved in anaplerosis/cataplerosis, reactive oxygen species buffering, and mitochondrial function, making it relevant to studies of metabolic remodeling and redox stress. Altered ME3 expression or dependency has been investigated in contexts where mitochondrial NADPH balance and oxidative metabolism contribute to disease-associated phenotypes, including cancer metabolism and mitochondrial dysfunction.
ME3 CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the ME3 gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the ME3 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 ME3 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 ME3 protein expression.
This CRISPR knockout system enables efficient generation of ME3-deficient cell models for investigation of ME3 signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting ME3 exon(s) critical for ME3 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 ME3 genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by ME3 CRISPR/Cas9 KO Plasmid (h) and ME3 CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the ME3 locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by ME3 HDR Plasmid (h) and ME3 HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by ME3 homology arms to support homology-directed repair at defined ME3 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.