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.
Paraplegin CRISPR/Cas9 KO Plasmid (h): sc-406299
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Paraplegin 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 Paraplegin 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: Paraplegin Antibody (C-5): sc-514393
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Paraplegin CRISPR/Cas9 KO Plasmid (h) | sc-406299 | 20 µg | $397.00 |
Overview
SPG7 encodes paraplegin, an ATP-dependent metalloprotease of the mitochondrial inner membrane m-AAA protease complex that supports protein quality control, proteolytic processing, and turnover of misfolded membrane and matrix-associated substrates. Paraplegin contributes to mitochondrial proteostasis, respiratory chain maintenance, and homeostatic responses to oxidative and proteotoxic stress through coordinated surveillance of inner membrane protein composition. Disruption of SPG7 perturbs mitochondrial function and is linked to neurodegenerative phenotypes, including hereditary spastic paraplegia, making it a useful target for studying axonal vulnerability and energy-dependent neuronal maintenance. SPG7-focused models also inform pathways involved in mitochondrial dynamics, unfolded protein responses, and stress-adaptive signaling.
Paraplegin CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the SPG7 gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the SPG7 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 SPG7 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 Paraplegin protein expression.
This CRISPR knockout system enables efficient generation of SPG7-deficient cell models for investigation of Paraplegin signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting SPG7 exon(s) critical for Paraplegin 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 SPG7 genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by Paraplegin CRISPR/Cas9 KO Plasmid (h) and Paraplegin CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the SPG7 locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by Paraplegin HDR Plasmid (h) and Paraplegin HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by SPG7 homology arms to support homology-directed repair at defined SPG7 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.