βB2-crystallin Double Nickase Plasmid (h): sc-403626-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- βB2-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
- βB2-crystallin Double Nickase Plasmid (h) and βB2-crystallin Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting CRYBB2. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: βB2-crystallin Antibody (B-12): sc-376006
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
βB2-crystallin Double Nickase Plasmid (h) | sc-403626-NIC | 20 µg | $410.00 | |||
βB2-crystallin Double Nickase Plasmid (h2) | sc-403626-NIC-2 | 20 µg | $410.00 |
CRYBB2 encodes human βB2-crystallin, a major structural protein of the ocular lens that supports lens fiber cell architecture and long-term transparency. As a member of the β/γ-crystallin superfamily, βB2-crystallin contributes to high-protein packing and refractive properties while maintaining proteostasis through chaperone-like interactions that limit aggregation under oxidative and age-related stress. Altered CRYBB2 expression or protein destabilization is linked to disrupted lens homeostasis and cataract-associated phenotypes, making it a useful model for studying protein stability, stress responses, and lens differentiation programs. βB2-crystallin also provides a tractable system for investigating how conserved crystallin networks respond to redox imbalance and proteotoxic stress in ocular and non-ocular cell contexts.
βB2-crystallin Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the CRYBB2 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within CRYBB2. 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 CRYBB2 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 CRYBB2-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.