FLG/Filaggrin Double Nickase Plasmid (h): sc-400644-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- FLG/Filaggrin 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
- FLG/Filaggrin Double Nickase Plasmid (h) and FLG/Filaggrin Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting FLG. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: FLG/Filaggrin Antibody (AKH1): sc-66192
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
FLG/Filaggrin Double Nickase Plasmid (h) | sc-400644-NIC | 20 µg | $410.00 | |||
FLG/Filaggrin Double Nickase Plasmid (h2) | sc-400644-NIC-2 | 20 µg | $410.00 |
FLG encodes filaggrin, a key structural protein produced as profilaggrin in differentiating keratinocytes and proteolytically processed into filaggrin monomers that aggregate keratin intermediate filaments. This function supports terminal epidermal differentiation, corneocyte compaction, and formation of the stratum corneum barrier, and its further breakdown products contribute to natural moisturizing factors that regulate hydration and skin surface pH. FLG activity is integrated with keratinization and cornified envelope assembly programs downstream of epidermal differentiation signaling networks. Genetic variation or reduced filaggrin expression is strongly associated with barrier dysfunction phenotypes and inflammatory skin disease susceptibility, making FLG a frequent target in studies of epithelial homeostasis and cutaneous inflammation mechanisms.
FLG/Filaggrin Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the FLG locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within FLG. 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 FLG 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 FLG-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.