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.
TIM Double Nickase Plasmid (h): sc-416503-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- TIM 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
- TIM Double Nickase Plasmid (h) and TIM Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting TPI1. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: TIM Antibody (H-11): sc-166785
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
TIM Double Nickase Plasmid (h) | sc-416503-NIC | 20 µg | $410.00 |
Human TPI1 encodes triosephosphate isomerase (TIM), a cytosolic glycolytic enzyme that catalyzes the interconversion of dihydroxyacetone phosphate and glyceraldehyde-3-phosphate. This step supports efficient glycolytic flux and links carbohydrate metabolism to downstream ATP production, redox balance, and biosynthetic pathways such as the pentose phosphate pathway. TPI1 activity contributes to cellular energy homeostasis and metabolic reprogramming observed in proliferative and stress-adapted states, making it relevant to studies of hypoxia responses and central carbon metabolism. Inherited TPI1 deficiency is associated with hemolytic anemia and neuromuscular dysfunction, and altered glycolytic control involving TPI1 is frequently investigated in cancer metabolism research.
TIM Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the TPI1 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within TPI1. 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 TPI1 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 TPI1-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.