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.
HPRT Double Nickase Plasmid (h): sc-417332-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- HPRT 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
- HPRT Double Nickase Plasmid (h) and HPRT Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting HPRT1. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: HPRT Antibody (F-1): sc-376938
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
HPRT Double Nickase Plasmid (h) | sc-417332-NIC | 20 µg | $410.00 | |||
HPRT Double Nickase Plasmid (h2) | sc-417332-NIC-2 | 20 µg | $410.00 |
HPRT1 encodes hypoxanthine-guanine phosphoribosyltransferase (HPRT), a key enzyme of the purine salvage pathway that recycles hypoxanthine and guanine to IMP and GMP, supporting nucleotide homeostasis and energy metabolism. HPRT activity contributes to cellular responses to nucleotide depletion and is central to selection schemes using purine analogs in cultured cells. Disruption of HPRT1 perturbs purine flux and can elevate uric acid production, linking the pathway to metabolic stress phenotypes. Genetic deficiency of HPRT1 is classically associated with Lesch–Nyhan spectrum disorders and provides a widely used model locus for studying mutation, selection, and DNA repair outcomes.
HPRT Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the HPRT1 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within HPRT1. 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 HPRT1 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 HPRT1-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.