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.
creatine kinase-M CRISPR/Cas9 KO Plasmid (h2): sc-401132-KO-2
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- creatine kinase-M CRISPR/Cas9 Knockout (KO) Plasmid (h2) 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 creatine kinase-M 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: creatine kinase-M Antibody (G-9): sc-365046
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
creatine kinase-M CRISPR/Cas9 KO Plasmid (h2) | sc-401132-KO-2 | 20 µg | $397.00 |
Overview
CKM encodes creatine kinase-M (CK-M), a cytosolic phosphotransferase that catalyzes reversible transfer of a phosphate group from phosphocreatine to ADP to regenerate ATP, thereby buffering cellular energy charge during fluctuating demand. Highly enriched in skeletal and cardiac muscle, CK-M supports ATP homeostasis coupled to glycolysis, oxidative phosphorylation, and phosphocreatine shuttle–mediated energy transport in myofibers. Perturbation of CKM activity impacts muscle contractility and metabolic stress responses and is frequently evaluated in studies of myopathies, muscle injury, and cardiometabolic remodeling. CKM also serves as a mechanistic node for investigating energetic adaptation, mitochondrial function, and redox-linked signaling in striated muscle models.
creatine kinase-M CRISPR/Cas9 KO Plasmid (h2) is a pool of plasmids designed for targeted disruption of the CKM gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the CKM 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 CKM 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 creatine kinase-M protein expression.
This CRISPR knockout system enables efficient generation of CKM-deficient cell models for investigation of creatine kinase-M signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting CKM exon(s) critical for creatine kinase-M 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 CKM genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by creatine kinase-M CRISPR/Cas9 KO Plasmid (h) and creatine kinase-M CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the CKM locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by creatine kinase-M HDR Plasmid (h) and creatine kinase-M HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by CKM homology arms to support homology-directed repair at defined CKM 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.