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.
Transketolase Double Nickase Plasmid (m): sc-423410-NIC
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Transketolase Double Nickase Plasmid (m) 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
- Transketolase Double Nickase Plasmid (m) and Transketolase Double Nickase Plasmid (m2) encode distinct paired gRNA designs targeting Tkt. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: Transketolase Antibody (H-7): sc-390179
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Transketolase Double Nickase Plasmid (m) | sc-423410-NIC | 20 µg | $410.00 |
Mouse Tkt encodes transketolase, a thiamine pyrophosphate–dependent enzyme that catalyzes reversible two-carbon transfer reactions in the non-oxidative branch of the pentose phosphate pathway. By interconverting sugar phosphates such as xylulose-5-phosphate, ribose-5-phosphate, and fructose-6-phosphate, transketolase links nucleotide biosynthesis with glycolytic flux and supports cellular redox balance indirectly through coordination with oxidative PPP activity. Tkt function is therefore relevant to proliferative metabolism, maintenance of NADPH-dependent antioxidant systems, and anabolic demands in rapidly dividing cells. Dysregulation of PPP enzymes, including transketolase, has been associated with altered metabolic homeostasis and oxidative stress phenotypes that are widely studied in metabolic and neurodegeneration-related research contexts.
Transketolase Double Nickase Plasmid (m) consists of a matched pair of plasmids engineered for high-specificity editing of the Tkt locus in mouse cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within Tkt. 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 Tkt 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 Tkt-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.