Pre-TCRα CRISPR/Cas9 KO Plasmid (m): sc-422473
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Pre-TCRα 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 Pre-TCRα 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
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Pre-TCRα CRISPR/Cas9 KO Plasmid (m) | sc-422473 | 20 µg | $397.00 |
Overview
Ptcra encodes the mouse pre–T cell receptor alpha (pre-TCRα), an essential component of the pre-TCR complex expressed in developing thymocytes. Pre-TCRα pairs with TCRβ to form a signaling receptor that promotes β-selection, driving proliferation, survival, and progression from the double-negative to double-positive stage. Pre-TCR signaling engages kinase cascades and transcriptional programs linked to thymocyte differentiation, including LCK/ZAP70-dependent pathways and downstream MAPK and PI3K–AKT signaling. Dysregulated PTCRA-associated checkpoints can perturb T cell development and repertoire formation, providing a mechanistic entry point for studying immune deficiency and T cell–driven pathophysiology in mouse models.
Pre-TCRα CRISPR/Cas9 KO Plasmid (m) is a pool of plasmids designed for targeted disruption of the Ptcra gene in mouse cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the Ptcra 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 Ptcra 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 Pre-TCRα protein expression.
This CRISPR knockout system enables efficient generation of Ptcra-deficient cell models for investigation of Pre-TCRα signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting Ptcra exon(s) critical for Pre-TCRα 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 Ptcra genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by Pre-TCRα CRISPR/Cas9 KO Plasmid (m) and Pre-TCRα CRISPR/Cas9 KO Plasmid (m2) target distinct sites within the Ptcra locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by Pre-TCRα HDR Plasmid (m) and Pre-TCRα HDR Plasmid (m2) contain a puromycin resistance cassette and an RFP reporter flanked by Ptcra homology arms to support homology-directed repair at defined Ptcra 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.