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.
ATF4 Double Nickase Plasmid (h): sc-400155-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- ATF4 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
- ATF4 Double Nickase Plasmid (h) and ATF4 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting ATF4. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: ATF4 Antibody (B-3): sc-390063
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
ATF4 Double Nickase Plasmid (h) | sc-400155-NIC | 20 µg | $410.00 | |||
ATF4 Double Nickase Plasmid (h2) | sc-400155-NIC-2 | 20 µg | $410.00 |
Activating transcription factor 4 (ATF4) is a stress-responsive bZIP transcription factor that coordinates adaptive gene expression programs during the integrated stress response (ISR). ATF4 is preferentially translated following eIF2α phosphorylation downstream of PERK, GCN2, PKR, and HRI, driving transcriptional networks that regulate amino acid metabolism, redox homeostasis, autophagy, and apoptosis through targets involved in glutathione synthesis, one-carbon metabolism, and proteostasis. It interfaces with ER stress and unfolded protein response signaling, including cooperation with CHOP/DDIT3, to shape cell-fate decisions under proteotoxic or nutrient stress. Dysregulated ATF4 activity has been implicated in cancer cell survival under hypoxia and nutrient limitation, neurodegeneration and proteostasis disorders, and metabolic and inflammatory states where stress-adaptive transcriptional control is altered.
ATF4 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the ATF4 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within ATF4. 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 ATF4 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 ATF4-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.