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.
LAT Double Nickase Plasmid (m): sc-421389-NIC
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- LAT Double Nickase Plasmid (m) 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
- LAT Double Nickase Plasmid (m) and LAT Double Nickase Plasmid (m2) encode distinct paired gRNA designs targeting Lat. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: LAT Antibody (11B.12): sc-53550
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
LAT Double Nickase Plasmid (m) | sc-421389-NIC | 20 µg | $410.00 |
Mouse LAT (linker for activation of T cells) encodes a transmembrane adaptor protein that is rapidly phosphorylated downstream of T cell receptor engagement, creating docking sites for SH2-domain signaling proteins. LAT nucleates the LAT signalosome to couple proximal kinases such as LCK and ZAP70 to PLCγ1 activation, calcium flux, Ras–MAPK signaling, and PI3K-dependent pathways that control T cell activation, differentiation, and cytokine production. Disruption of LAT signaling alters immune synapse formation and T cell selection, and LAT pathway dysfunction is associated with immune dysregulation and lymphoproliferative phenotypes in experimental models. As a central hub in adaptive immune signaling, LAT is widely studied in mechanisms of T cell tolerance, inflammation, and immune-mediated disease biology.
LAT Double Nickase Plasmid (m) consists of a matched pair of plasmids engineered for high-specificity editing of the Lat locus in mouse cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within Lat. 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 Lat 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 Lat-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.