LysRS Double Nickase Plasmid (h): sc-404110-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- LysRS 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
- LysRS Double Nickase Plasmid (h) and LysRS Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting KARS. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: LysRS Antibody (A-10): sc-271331
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
LysRS Double Nickase Plasmid (h) | sc-404110-NIC | 20 µg | $410.00 | |||
LysRS Double Nickase Plasmid (h2) | sc-404110-NIC-2 | 20 µg | $410.00 |
KARS encodes human lysyl-tRNA synthetase (LysRS), a class II aminoacyl-tRNA synthetase that ligates lysine to its cognate tRNALys, ensuring translational fidelity and supporting proteome homeostasis. Beyond its canonical role in cytosolic protein synthesis, LysRS has been implicated in signaling-linked processes through regulated localization and protein–protein interactions that connect translation with cellular stress responses. Perturbation of aminoacyl-tRNA synthetase function can alter proteostasis, integrated stress signaling, and cell-state programs that are frequently interrogated in models of neurodevelopment, neurodegeneration, and cancer biology. Dysregulated KARS activity or expression is also relevant to mitochondrial–cytosolic coordination of gene expression and metabolic adaptation, making it a useful node for studying translation-centric mechanisms.
LysRS Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the KARS locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within KARS. 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 KARS 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 KARS-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.