GADD 34 Double Nickase Plasmid (h): sc-400595-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- GADD 34 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
- GADD 34 Double Nickase Plasmid (h) and GADD 34 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting PPP1R15A. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: GADD 34 Antibody (B-10): sc-373815
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
GADD 34 Double Nickase Plasmid (h) | sc-400595-NIC | 20 µg | $410.00 | |||
GADD 34 Double Nickase Plasmid (h2) | sc-400595-NIC-2 | 20 µg | $410.00 |
PPP1R15A encodes GADD34, an inducible regulatory subunit of protein phosphatase 1 that promotes dephosphorylation of eIF2α to terminate the integrated stress response and restore translation after cellular stress. GADD34 is upregulated downstream of ER stress and DNA damage signaling and modulates proteostasis, apoptosis susceptibility, and feedback control of the unfolded protein response. Through control of eIF2α phosphorylation dynamics, PPP1R15A influences ATF4/CHOP-associated transcriptional programs and stress-adaptive remodeling of gene expression. Dysregulation of this axis has been linked to pathological states characterized by chronic ER stress and altered stress tolerance, supporting its study in neurodegeneration, metabolic stress, and cancer cell biology.
GADD 34 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the PPP1R15A locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within PPP1R15A. 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 PPP1R15A 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 PPP1R15A-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.