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.
MCAT CRISPR/Cas9 KO Plasmid (h): sc-404338
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- MCAT 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 MCAT 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: MCAT Antibody (E-11): sc-390858
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
MCAT CRISPR/Cas9 KO Plasmid (h) | sc-404338 | 20 µg | $397.00 |
Overview
MCAT (malonyl-CoA:ACP transacylase) is a mitochondrial enzyme required for mitochondrial fatty acid synthesis (mtFAS) by transferring malonyl groups to acyl carrier protein, enabling elongation of acyl chains used in lipoic acid production. Through its role in generating lipoate, MCAT supports lipoylation of key dehydrogenase complexes such as pyruvate dehydrogenase and α-ketoglutarate dehydrogenase, linking mtFAS to central carbon metabolism, redox homeostasis, and mitochondrial energy flux. Perturbation of MCAT activity can influence mitochondrial respiration and metabolite utilization, with downstream effects on oxidative stress and metabolic reprogramming. These processes are relevant to mechanistic studies of mitochondrial dysfunction and metabolism-associated disease biology, including contexts where lipoylation status and TCA-linked enzyme function are altered.
MCAT CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the MCAT gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the MCAT 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 MCAT 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 MCAT protein expression.
This CRISPR knockout system enables efficient generation of MCAT-deficient cell models for investigation of MCAT signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting MCAT exon(s) critical for MCAT 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 MCAT genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by MCAT CRISPR/Cas9 KO Plasmid (h) and MCAT CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the MCAT locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by MCAT HDR Plasmid (h) and MCAT HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by MCAT homology arms to support homology-directed repair at defined MCAT 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.