βB1-crystallin Double Nickase Plasmid (h): sc-403361-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- βB1-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
- βB1-crystallin Double Nickase Plasmid (h) and βB1-crystallin Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting CRYBB1. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: βB1-crystallin Antibody (A-8): sc-374496
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
βB1-crystallin Double Nickase Plasmid (h) | sc-403361-NIC | 20 µg | $410.00 | |||
βB1-crystallin Double Nickase Plasmid (h2) | sc-403361-NIC-2 | 20 µg | $410.00 |
CRYBB1 encodes human βB1-crystallin, a major structural protein of the eye lens that contributes to the high refractive index and long-term transparency required for vision. βB1-crystallin participates in the crystallin protein network by forming stable oligomers with other β/γ-crystallins, supporting lens fiber cell architecture and protein homeostasis. Disruption of crystallin assembly or increased susceptibility to aggregation can compromise lens clarity, making CRYBB1 a key locus for studying proteostasis, stress responses, and age-related changes in lens biology. Variants affecting βB1-crystallin stability and solubility have been linked to inherited cataract phenotypes, providing a genetic context for mechanistic studies in ocular models.
βB1-crystallin Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the CRYBB1 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within CRYBB1. 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 CRYBB1 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 CRYBB1-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.