GS1 Double Nickase Plasmid (h): sc-405996-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- GS1 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
- GS1 Double Nickase Plasmid (h) and GS1 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting PUDP. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: GS1 Antibody (B-4): sc-166043
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
GS1 Double Nickase Plasmid (h) | sc-405996-NIC | 20 µg | $410.00 | |||
GS1 Double Nickase Plasmid (h2) | sc-405996-NIC-2 | 20 µg | $410.00 |
Human PUDP encodes pseudouridine-5′-phosphatase (GS1), an enzyme in the pyrimidine nucleoside salvage network that dephosphorylates pseudouridine monophosphate to pseudouridine, supporting nucleotide homeostasis and RNA turnover. This activity links RNA modification catabolism to central carbon metabolism by channeling ribose-1-phosphate and related intermediates into broader metabolic pathways. Altered PUDP/GS1 function can perturb cellular nucleotide balance and metabolic state, processes frequently coupled to proliferative stress and genome maintenance programs. As a result, PUDP is of interest for mechanistic studies of RNA modification metabolism, stress adaptation, and pathway rewiring in disease-relevant cellular models.
GS1 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the PUDP locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within PUDP. 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 PUDP 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 PUDP-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.