DDIT3/CHOP/GADD153 Double Nickase Plasmid (h): sc-400051-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- DDIT3/CHOP/GADD153 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
- DDIT3/CHOP/GADD153 Double Nickase Plasmid (h) and DDIT3/CHOP/GADD153 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting DDIT3. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: DDIT3/GADD153/CHOP Antibody (B-3): sc-7351
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
DDIT3/CHOP/GADD153 Double Nickase Plasmid (h) | sc-400051-NIC | 20 µg | $410.00 | |||
DDIT3/CHOP/GADD153 Double Nickase Plasmid (h2) | sc-400051-NIC-2 | 20 µg | $410.00 |
DDIT3 encodes the transcription factor CHOP (GADD153), a stress-inducible bZIP protein that heterodimerizes with C/EBP family members to reprogram gene expression during cellular stress. CHOP is a central effector of the unfolded protein response (UPR), integrating PERK–eIF2α–ATF4 signaling to regulate apoptosis, redox control, and proteostasis under endoplasmic reticulum stress. Beyond ER stress, DDIT3 participates in integrated stress response pathways linked to nutrient deprivation, oxidative stress, and inflammatory cues, influencing cell fate decisions and differentiation programs. Dysregulated CHOP activity is implicated in metabolic dysfunction, neurodegeneration models, inflammatory tissue injury, and oncogenic contexts including the FUS-DDIT3 fusion associated with myxoid liposarcoma, making DDIT3 a key node for mechanistic studies of stress-adaptive transcriptional networks.
DDIT3/CHOP/GADD153 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the DDIT3 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within DDIT3. 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 DDIT3 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 DDIT3-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.