CD28 CRISPR/Cas9 KO Plasmid (m): sc-419541
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- CD28 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 CD28 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: CD28 Antibody (3H1179): sc-70612
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
CD28 CRISPR/Cas9 KO Plasmid (m) | sc-419541 | 20 µg | $397.00 |
Overview
Cd28 encodes CD28, a key costimulatory receptor on T cells that augments TCR signaling to support full activation, IL-2 production, proliferation, and survival. CD28 engagement by CD80/CD86 on antigen-presenting cells activates PI3K–AKT–mTOR and NF-κB/AP-1 transcriptional programs, shaping effector differentiation and metabolic reprogramming. By setting activation thresholds and promoting regulatory T cell homeostasis, CD28 influences peripheral tolerance, chronic inflammation, and immune dysregulation phenotypes observed in mouse models of autoimmunity and infection. Altered CD28 signaling is also relevant to tumor immune evasion and immunopathology studies that interrogate checkpoint and costimulatory networks.
CD28 CRISPR/Cas9 KO Plasmid (m) is a pool of plasmids designed for targeted disruption of the Cd28 gene in mouse cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the Cd28 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 Cd28 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 CD28 protein expression.
This CRISPR knockout system enables efficient generation of Cd28-deficient cell models for investigation of CD28 signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting Cd28 exon(s) critical for CD28 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 Cd28 genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by CD28 CRISPR/Cas9 KO Plasmid (m) and CD28 CRISPR/Cas9 KO Plasmid (m2) target distinct sites within the Cd28 locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by CD28 HDR Plasmid (m) and CD28 HDR Plasmid (m2) contain a puromycin resistance cassette and an RFP reporter flanked by Cd28 homology arms to support homology-directed repair at defined Cd28 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.