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-416991-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- DHFRL1 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
- DHFRL1 Double Nickase Plasmid (h) and DHFRL1 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting DHFR2. One or both designs may be available
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
DHFR Double Nickase Plasmid (h) | sc-416991-NIC | 20 µg | $410.00 | |||
DHFRL1 Double Nickase Plasmid (h2) | sc-416991-NIC-2 | 20 µg | $410.00 |
Human DHFR2 encodes dihydrofolate reductase (DHFR), an NADPH-dependent enzyme that catalyzes reduction of dihydrofolate to tetrahydrofolate, sustaining one-carbon metabolism required for de novo thymidylate and purine biosynthesis. Through its role in folate cycling, DHFR supports DNA replication and repair, cell-cycle progression, and cellular methylation capacity via linkage to methionine/S-adenosylmethionine metabolism. Perturbation of DHFR activity influences nucleotide pool balance and genome stability, processes frequently examined in proliferative stress and metabolic remodeling contexts. DHFR biology is therefore widely leveraged to study folate-dependent biosynthetic pathways and cellular responses to nucleotide depletion and replication stress.
DHFRL1 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the DHFR2 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within DHFR2. 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 DHFR2 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 DHFR2-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.