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.
DHFR Double Nickase Plasmid (h): sc-417022-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- DHFR 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
- DHFR Double Nickase Plasmid (h) and DHFR Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting DHFR. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: DHFR Antibody (F-8): sc-74594
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
DHFR Double Nickase Plasmid (h) | sc-417022-NIC | 20 µg | $410.00 | |||
DHFR Double Nickase Plasmid (h2) | sc-417022-NIC-2 | 20 µg | $410.00 |
Human DHFR (dihydrofolate reductase) catalyzes the NADPH-dependent reduction of dihydrofolate to tetrahydrofolate, sustaining one-carbon metabolism required for de novo purine and thymidylate biosynthesis. Through its central role in nucleotide production, DHFR supports DNA replication and repair, S-phase progression, and proliferation, linking folate metabolism to genome maintenance. DHFR activity is tightly connected to metabolic stress responses and cellular sensitivity to antifolate pressure, making it a widely used marker in studies of metabolic adaptation and selection. Dysregulated folate pathway flux and DHFR expression or copy-number changes have been associated with proliferative phenotypes and drug response variability in multiple disease contexts.
DHFR Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the DHFR locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within DHFR. 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 DHFR 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 DHFR-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.