ENT4 Double Nickase Plasmid (h): sc-405115-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- ENT4 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
- ENT4 Double Nickase Plasmid (h) and ENT4 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting SLC29A4. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: ENT4 Antibody (SA-18): sc-101295
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
ENT4 Double Nickase Plasmid (h) | sc-405115-NIC | 20 µg | $410.00 | |||
ENT4 Double Nickase Plasmid (h2) | sc-405115-NIC-2 | 20 µg | $410.00 |
Human SLC29A4 encodes equilibrative nucleoside transporter 4 (ENT4), a member of the SLC29 family that mediates bidirectional transport of nucleosides and related organic cations across cellular membranes according to concentration gradients. ENT4 activity contributes to nucleoside salvage and purine metabolism, influencing intracellular nucleotide pools and thereby coupling membrane transport to DNA/RNA synthesis and broader metabolic homeostasis. Transport by ENT4 is sensitive to extracellular conditions and can intersect with cellular stress responses that reshape nucleotide demand and signaling. Altered SLC29A4/ENT4 function has been investigated in the context of dysregulated nucleoside handling and metabolite-driven phenotypes relevant to neurological and metabolic disease research.
ENT4 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the SLC29A4 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within SLC29A4. 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 SLC29A4 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 SLC29A4-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.