Sam50 CRISPR/Cas9 KO Plasmid (m): sc-427129
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Sam50 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 Sam50 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: Sam50 Antibody (SQ-7): sc-100493
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Sam50 CRISPR/Cas9 KO Plasmid (m) | sc-427129 | 20 µg | $397.00 |
Overview
Samm50 encodes Sam50, an essential β-barrel protein of the mitochondrial outer membrane that forms the core of the sorting and assembly machinery (SAM) complex. Sam50 coordinates the biogenesis and membrane insertion of β-barrel proteins and functionally interfaces with mitochondrial protein import systems, supporting outer membrane organization and mitochondrial homeostasis. Through its role in maintaining mitochondrial architecture, protein import, and respiration-linked processes, altered SAM complex function is relevant to studies of mitochondrial dysfunction, stress signaling, and pathways associated with neurodegeneration and metabolic disease phenotypes in model systems. In mouse cells, Samm50 perturbation is also used to investigate mitochondrial quality control, including crosstalk with mitophagy and intrinsic apoptosis signaling.
Sam50 CRISPR/Cas9 KO Plasmid (m) is a pool of plasmids designed for targeted disruption of the Samm50 gene in mouse cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the Samm50 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 Samm50 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 Sam50 protein expression.
This CRISPR knockout system enables efficient generation of Samm50-deficient cell models for investigation of Sam50 signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting Samm50 exon(s) critical for Sam50 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 Samm50 genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by Sam50 CRISPR/Cas9 KO Plasmid (m) and Sam50 CRISPR/Cas9 KO Plasmid (m2) target distinct sites within the Samm50 locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by Sam50 HDR Plasmid (m) and Sam50 HDR Plasmid (m2) contain a puromycin resistance cassette and an RFP reporter flanked by Samm50 homology arms to support homology-directed repair at defined Samm50 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.