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.
Gads CRISPR/Cas9 KO Plasmid (m): sc-421689
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Gads 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 Gads 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: Gads Antibody (UW40): sc-73652
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Gads CRISPR/Cas9 KO Plasmid (m) | sc-421689 | 20 µg | $397.00 |
Overview
Grap2 encodes the adaptor protein Gads, a hematopoietic signaling component that links activated immune receptors to downstream effector pathways. In T cells, Gads couples phosphorylated LAT to SLP-76 to assemble signaling microclusters that propagate proximal T cell receptor signaling, facilitating PLCγ1 activation, calcium flux, MAPK signaling, and transcriptional programs controlling cytokine production and proliferation. This adaptor also participates in signaling from other immunoreceptors in lymphoid lineages, influencing cell activation thresholds and immune synapse organization. Dysregulated Gads-dependent signaling networks are frequently examined in models of immune dysfunction, inflammatory phenotypes, and hematopoietic signaling abnormalities.
Gads CRISPR/Cas9 KO Plasmid (m) is a pool of plasmids designed for targeted disruption of the Grap2 gene in mouse cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the Grap2 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 Grap2 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 Gads protein expression.
This CRISPR knockout system enables efficient generation of Grap2-deficient cell models for investigation of Gads signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting Grap2 exon(s) critical for Gads 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 Grap2 genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by Gads CRISPR/Cas9 KO Plasmid (m) and Gads CRISPR/Cas9 KO Plasmid (m2) target distinct sites within the Grap2 locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by Gads HDR Plasmid (m) and Gads HDR Plasmid (m2) contain a puromycin resistance cassette and an RFP reporter flanked by Grap2 homology arms to support homology-directed repair at defined Grap2 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.