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.
CD28 Double Nickase Plasmid (h): sc-401525-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- CD28 Double Nickase Plasmid (h) consists of a pair of plasmids each encoding a D10A mutated Cas9 nuclease and a target-specific 20 nt guide RNA (gRNA) designed to knockout gene expression with greater specificity than its CRISPR/Cas9 KO counterpart
- Paired gRNA sequences are offset by approximately 20 bp to allow for specific Cas9-mediated double nicking of the genomic DNA, which mimics a DSB
- One plasmid in the pair contains a puromycin-resistance gene for selection; the other plasmid in the pair contains a GFP marker to visually confirm transfection
- CD28 Double Nickase Plasmid (h) and CD28 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting CD28. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: CD28 Antibody (CD28.2): sc-19655
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
CD28 Double Nickase Plasmid (h) | sc-401525-NIC | 20 µg | $410.00 | |||
CD28 Double Nickase Plasmid (h2) | sc-401525-NIC-2 | 20 µg | $410.00 |
CD28 encodes a co-stimulatory receptor expressed on T cells that cooperates with T cell receptor signaling to promote activation, proliferation, cytokine production, and survival. Engagement of CD28 by CD80/CD86 on antigen-presenting cells amplifies PI3K–AKT–mTOR signaling, supports NF-κB and MAPK pathway activation, and enhances IL-2 transcription, shaping effector differentiation and memory formation. CD28 activity helps balance immune tolerance and responsiveness, with altered signaling linked to dysregulated T cell activation and immune-mediated pathology. Because CD28 is central to co-stimulatory checkpoint control, it is widely studied in contexts such as autoimmunity, chronic inflammation, infection, and tumor immunology.
CD28 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the CD28 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within CD28. When directed to adjacent sites on opposite DNA strands, the two nickases generate offset single-strand nicks that together produce a staggered double-strand break, requiring coordinated on-target activity from both guides. The resulting DNA break is resolved by endogenous cellular repair pathways, most commonly through non-homologous end joining (NHEJ), leading to insertions or deletions that disrupt CD28 function. By requiring dual sgRNA engagement at the target locus, the double nicking approach enhances editing specificity and provides a complementary CRISPR strategy for applications where additional control over targeting precision is desired.
To support efficient identification of edited cells, one plasmid encodes GFP for fluorescent visualization of transfected populations, while the companion plasmid carries a puromycin resistance gene for antibiotic selection. Together, these features support efficient enrichment of co-transfected populations and simplify the validation of CD28-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.