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.
GFAP Double Nickase Plasmid (h): sc-400130-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- GFAP 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
- GFAP Double Nickase Plasmid (h) and GFAP Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting GFAP. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: GFAP Antibody (2E1): sc-33673
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
GFAP Double Nickase Plasmid (h) | sc-400130-NIC | 20 µg | $410.00 | |||
GFAP Double Nickase Plasmid (h2) | sc-400130-NIC-2 | 20 µg | $410.00 |
Glial fibrillary acidic protein (GFAP) is an intermediate filament protein that serves as a principal structural component of the astrocytic cytoskeleton and is widely used as a marker of astrocyte differentiation and reactive gliosis. By organizing filament networks and interacting with cytoskeletal regulators, GFAP contributes to astrocyte morphology, mechanical stability, and the cellular response to injury and inflammatory cues. GFAP dynamics intersect with processes such as cytoskeletal remodeling, cell migration, and stress signaling that shape glial scar formation and neuroimmune interactions. Altered GFAP expression or aggregation is associated with astrocyte dysfunction in neurological and neurodegenerative contexts, including Alexander disease and broader CNS injury responses.
GFAP Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the GFAP locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within GFAP. 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 GFAP 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 GFAP-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.