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.
GlyRS Double Nickase Plasmid (h): sc-404105-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- GlyRS 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
- GlyRS Double Nickase Plasmid (h) and GlyRS Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting GARS. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: GlyRS Antibody (D-10): sc-365311
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
GlyRS Double Nickase Plasmid (h) | sc-404105-NIC | 20 µg | $410.00 | |||
GlyRS Double Nickase Plasmid (h2) | sc-404105-NIC-2 | 20 µg | $410.00 |
GARS encodes human glycyl-tRNA synthetase (GlyRS), a cytosolic aminoacyl-tRNA synthetase that charges tRNAGly with glycine, supplying aminoacylated tRNAs required for ribosomal translation. By maintaining translational fidelity and proteostasis, GlyRS supports core processes including protein synthesis, stress responses, and energy-demanding cellular states where high translational throughput is required. Dysregulation or mutation of GARS has been linked to peripheral neuropathy phenotypes, highlighting the sensitivity of neuronal and axonal systems to altered tRNA charging and protein homeostasis. As a conserved housekeeping enzyme with tissue-specific vulnerability, GlyRS is frequently studied in models of neurodegeneration, proteotoxic stress, and translation-dependent signaling.
GlyRS Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the GARS locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within GARS. 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 GARS 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 GARS-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.