PBGD Double Nickase Plasmid (h): sc-404924-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- PBGD 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
- PBGD Double Nickase Plasmid (h) and PBGD Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting HMBS. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: PBGD Antibody (E-9): sc-166743
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
PBGD Double Nickase Plasmid (h) | sc-404924-NIC | 20 µg | $410.00 | |||
PBGD Double Nickase Plasmid (h2) | sc-404924-NIC-2 | 20 µg | $410.00 |
Human HMBS encodes porphobilinogen deaminase (PBGD), a cytosolic enzyme that catalyzes the polymerization of porphobilinogen into hydroxymethylbilane during heme biosynthesis. This step supports production of heme required for mitochondrial respiration, cytochrome function, and oxygen handling in erythroid and non-erythroid tissues. HMBS activity integrates with porphyrin metabolism and cellular redox homeostasis, linking heme availability to oxidative stress responses and metabolic adaptation. Genetic or functional perturbation of HMBS is associated with defects in porphyrin pathway flux and is relevant to studies of acute hepatic porphyrias and erythroid heme regulation.
PBGD Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the HMBS locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within HMBS. 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 HMBS 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 HMBS-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.