TorsinA CRISPR/Cas9 KO Plasmid (m): sc-424414
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- TorsinA 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 TorsinA 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: TorsinA Antibody (D-7): sc-373915
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
TorsinA CRISPR/Cas9 KO Plasmid (m) | sc-424414 | 20 µg | $397.00 |
Overview
Tor1a encodes torsinA, an AAA+ ATPase that localizes to the endoplasmic reticulum and nuclear envelope, where it contributes to nuclear envelope integrity, proteostasis, and membrane-associated remodeling through interactions with LAP1 and LULL1. In mouse cells, torsinA activity is linked to regulation of nucleocytoplasmic transport, endoplasmic reticulum stress responses, and quality control of misfolded proteins. Disruption of TOR1A function is associated with neuronal dysfunction and dystonia-related phenotypes, making it relevant for studies of motor circuit biology and cellular stress adaptation. Tor1a is also used as a model locus for investigating how nuclear envelope perturbations influence cytoskeletal coupling and intracellular trafficking.
TorsinA CRISPR/Cas9 KO Plasmid (m) is a pool of plasmids designed for targeted disruption of the Tor1a gene in mouse cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the Tor1a 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 Tor1a 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 TorsinA protein expression.
This CRISPR knockout system enables efficient generation of Tor1a-deficient cell models for investigation of TorsinA signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting Tor1a exon(s) critical for TorsinA 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 Tor1a genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by TorsinA CRISPR/Cas9 KO Plasmid (m) and TorsinA CRISPR/Cas9 KO Plasmid (m2) target distinct sites within the Tor1a locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by TorsinA HDR Plasmid (m) and TorsinA HDR Plasmid (m2) contain a puromycin resistance cassette and an RFP reporter flanked by Tor1a homology arms to support homology-directed repair at defined Tor1a 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.