DAP-5 Double Nickase Plasmid (h): sc-405901-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- DAP-5 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
- DAP-5 Double Nickase Plasmid (h) and DAP-5 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting EIF4G2. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: DAP-5 Antibody (F-2): sc-137011
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
DAP-5 Double Nickase Plasmid (h) | sc-405901-NIC | 20 µg | $410.00 | |||
DAP-5 Double Nickase Plasmid (h2) | sc-405901-NIC-2 | 20 µg | $410.00 |
EIF4G2 encodes DAP-5 (also known as eIF4G2/NAT1), a noncanonical translation initiation scaffold that supports cap-independent and IRES-dependent mRNA translation, particularly under conditions where eIF4F activity is limited. DAP-5 contributes to translational control during cellular stress responses, apoptosis, and differentiation by coordinating interactions with initiation factors and selective mRNA recruitment. Through these mechanisms, EIF4G2 influences proteostasis and signaling programs that reshape gene expression at the level of translation. Dysregulation of EIF4G2-linked translational reprogramming has been investigated in contexts including tumor cell survival, neurodevelopmental processes, and stress-adaptive phenotypes, making it a useful node for mechanistic studies of RNA regulation.
DAP-5 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the EIF4G2 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within EIF4G2. 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 EIF4G2 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 EIF4G2-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.