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.
MCAD CRISPR/Cas9 KO Plasmid (m): sc-418938
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- MCAD 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 MCAD 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: MCAD Antibody (E-8): sc-271931
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
MCAD CRISPR/Cas9 KO Plasmid (m) | sc-418938 | 20 µg | $397.00 |
Overview
Acadm encodes medium-chain acyl-CoA dehydrogenase (MCAD), a mitochondrial flavoprotein that catalyzes the first dehydrogenation step in β-oxidation of medium-chain fatty acids, linking lipid catabolism to cellular ATP production and redox balance. MCAD activity supports metabolic flexibility during fasting and high-energy demand by fueling the TCA cycle and oxidative phosphorylation through acetyl-CoA and electron transfer via ETF/ETFDH. Disruption of Acadm perturbs mitochondrial fatty acid oxidation, alters acylcarnitine profiles, and can promote secondary effects on oxidative stress responses and lipid signaling networks. MCAD deficiency is a well-established inborn error of metabolism associated with impaired fatty acid utilization, making Acadm a relevant node for studying mitochondrial metabolic homeostasis in mouse models.
MCAD CRISPR/Cas9 KO Plasmid (m) is a pool of plasmids designed for targeted disruption of the Acadm gene in mouse cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the Acadm 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 Acadm 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 MCAD protein expression.
This CRISPR knockout system enables efficient generation of Acadm-deficient cell models for investigation of MCAD signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting Acadm exon(s) critical for MCAD 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 Acadm genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by MCAD CRISPR/Cas9 KO Plasmid (m) and MCAD CRISPR/Cas9 KO Plasmid (m2) target distinct sites within the Acadm locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by MCAD HDR Plasmid (m) and MCAD HDR Plasmid (m2) contain a puromycin resistance cassette and an RFP reporter flanked by Acadm homology arms to support homology-directed repair at defined Acadm 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.