AlaRS Double Nickase Plasmid (h): sc-404252-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- AlaRS 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
- AlaRS Double Nickase Plasmid (h) and AlaRS Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting AARS. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: AlaRS Antibody (A-6): sc-165990
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
AlaRS Double Nickase Plasmid (h) | sc-404252-NIC | 20 µg | $410.00 | |||
AlaRS Double Nickase Plasmid (h2) | sc-404252-NIC-2 | 20 µg | $410.00 |
AARS encodes human alanyl-tRNA synthetase (AlaRS), a cytosolic aminoacyl-tRNA synthetase that ligates alanine to its cognate tRNA, ensuring accurate decoding during translation. By maintaining tRNA charging fidelity, AlaRS supports global proteostasis and couples to cellular stress responses that monitor translational quality control. Perturbation of aminoacyl-tRNA synthetase activity can disrupt protein synthesis homeostasis and has been associated with neurodevelopmental and neurodegenerative phenotypes, highlighting AARS as a useful node for studying how translation-linked pathways influence cellular fitness. AARS function is also relevant to investigations of tRNA biology, ribosome dynamics, and downstream signaling changes triggered by impaired aminoacylation.
AlaRS Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the AARS locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within AARS. 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 AARS 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 AARS-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.