αA-crystallin Double Nickase Plasmid (h): sc-402047-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- αA-crystallin 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
- αA-crystallin Double Nickase Plasmid (h) and αA-crystallin Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting CRYAA. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: αA-crystallin Antibody (B-2): sc-28306
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
αA-crystallin Double Nickase Plasmid (h) | sc-402047-NIC | 20 µg | $410.00 | |||
αA-crystallin Double Nickase Plasmid (h2) | sc-402047-NIC-2 | 20 µg | $410.00 |
CRYAA encodes human αA-crystallin, a small heat shock protein that functions as an ATP-independent molecular chaperone to stabilize partially unfolded proteins and limit aggregation under stress. In the ocular lens, αA-crystallin supports proteostasis and contributes to lens transparency by buffering oxidative, thermal, and UV-induced damage, while also modulating cytoskeletal organization and apoptotic signaling in multiple cell types. The protein participates in stress-response networks linked to unfolded protein handling and maintenance of long-lived protein assemblies. Dysregulation or mutation of CRYAA is associated with cataractogenesis and has been studied in the context of proteotoxic stress responses relevant to neurodegenerative and myopathic phenotypes.
αA-crystallin Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the CRYAA locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within CRYAA. 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 CRYAA 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 CRYAA-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.