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.
CAR Double Nickase Plasmid (h): sc-418548-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- CAR 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
- CAR Double Nickase Plasmid (h) and CAR Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting CXADR. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: CAR Antibody (E-1): sc-373791
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
CAR Double Nickase Plasmid (h) | sc-418548-NIC | 20 µg | $410.00 | |||
CAR Double Nickase Plasmid (h2) | sc-418548-NIC-2 | 20 µg | $410.00 |
CXADR encodes the Coxsackievirus and adenovirus receptor (CAR), an immunoglobulin superfamily transmembrane protein that localizes to epithelial tight junctions and contributes to cell–cell adhesion and barrier organization. CAR participates in junctional assembly and cytoskeletal coupling through interactions with scaffolding proteins such as MAGI and PDZ domain–containing partners, influencing epithelial polarity and tissue architecture. Altered CXADR expression and CAR localization have been associated with changes in epithelial integrity and signaling programs relevant to cancer biology, myocardial remodeling, and susceptibility to viral entry, supporting its use as a mechanistic node in host–pathogen and junctional biology studies.
CAR Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the CXADR locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within CXADR. 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 CXADR 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 CXADR-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.