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.
Zap-70 CRISPR/Cas9 KO Plasmid (m): sc-423755
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Zap-70 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 Zap-70 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: ZAP-70 Antibody (1E7.2): sc-32760
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Zap-70 CRISPR/Cas9 KO Plasmid (m) | sc-423755 | 20 µg | $397.00 |
Overview
Zap70 encodes the cytosolic tyrosine kinase ZAP-70, a core signaling mediator downstream of the T cell receptor (TCR) and CD3 complex that links antigen recognition to intracellular phosphorylation cascades. Upon TCR engagement, ZAP-70 is recruited to phosphorylated ITAMs on CD3 chains and propagates signaling through LAT/SLP-76 scaffolding, driving PLCγ1 activation, calcium flux, MAPK signaling, and NFAT/NF-κB/AP-1 transcriptional programs. In mouse immune biology, Zap70 is essential for thymocyte selection, T cell maturation, and effector responses, making it central to studies of adaptive immunity and lymphocyte signaling networks. Dysregulated ZAP-70 pathway activity is relevant to models of immunodeficiency, autoimmunity, and lymphoid malignancy biology, where altered signaling thresholds influence development and activation.
Zap-70 CRISPR/Cas9 KO Plasmid (m) is a pool of plasmids designed for targeted disruption of the Zap70 gene in mouse cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the Zap70 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 Zap70 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 Zap-70 protein expression.
This CRISPR knockout system enables efficient generation of Zap70-deficient cell models for investigation of Zap-70 signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting Zap70 exon(s) critical for Zap-70 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 Zap70 genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by Zap-70 CRISPR/Cas9 KO Plasmid (m) and Zap-70 CRISPR/Cas9 KO Plasmid (m2) target distinct sites within the Zap70 locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by Zap-70 HDR Plasmid (m) and Zap-70 HDR Plasmid (m2) contain a puromycin resistance cassette and an RFP reporter flanked by Zap70 homology arms to support homology-directed repair at defined Zap70 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.