α-actinin-3 CRISPR/Cas9 KO Plasmid (m): sc-418972
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- α-actinin-3 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 α-actinin-3 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: α-actinin Antibody (H-2): sc-17829
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
α-actinin-3 CRISPR/Cas9 KO Plasmid (m) | sc-418972 | 20 µg | $397.00 |
Overview
Actn3 encodes α-actinin-3, a sarcomeric actin-binding protein enriched at the Z-disc of fast-twitch skeletal muscle fibers where it crosslinks and stabilizes thin filaments during contraction. By organizing the contractile apparatus and contributing to sarcomere mechanics, α-actinin-3 influences muscle performance traits and fiber-type properties linked to glycolytic metabolism and mechanical load response. Loss of α-actinin-3 function is associated with altered muscle strength and endurance phenotypes and has been investigated in the context of myopathies, muscle wasting, and adaptation to exercise or disuse. In mouse models, Actn3 provides a tractable system to study structural myofibrillar networks and pathways regulating contractility, muscle remodeling, and stress responses.
α-actinin-3 CRISPR/Cas9 KO Plasmid (m) is a pool of plasmids designed for targeted disruption of the Actn3 gene in mouse cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the Actn3 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 Actn3 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 α-actinin-3 protein expression.
This CRISPR knockout system enables efficient generation of Actn3-deficient cell models for investigation of α-actinin-3 signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting Actn3 exon(s) critical for α-actinin-3 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 Actn3 genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by α-actinin-3 CRISPR/Cas9 KO Plasmid (m) and α-actinin-3 CRISPR/Cas9 KO Plasmid (m2) target distinct sites within the Actn3 locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by α-actinin-3 HDR Plasmid (m) and α-actinin-3 HDR Plasmid (m2) contain a puromycin resistance cassette and an RFP reporter flanked by Actn3 homology arms to support homology-directed repair at defined Actn3 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.