PTRF Double Nickase Plasmid (h): sc-404669-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- PTRF 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
- PTRF Double Nickase Plasmid (h) and PTRF Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting CAVIN1. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: PTRF Antibody (4a): sc-517589
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
PTRF Double Nickase Plasmid (h) | sc-404669-NIC | 20 µg | $410.00 |
CAVIN1 (PTRF) encodes polymerase I and transcript release factor, an essential caveolae-associated protein that cooperates with caveolins to drive caveola biogenesis and stabilize plasma membrane caveolar domains. PTRF supports membrane organization, mechanoprotection, and lipid homeostasis by regulating caveolar endocytosis and caveola-dependent signaling, influencing processes such as insulin receptor signaling and cellular responses to membrane tension. Loss or dysfunction of CAVIN1 perturbs caveola formation and alters trafficking and signal compartmentalization, affecting adipocyte biology, muscle physiology, and vascular cell function. Genetic disruption of CAVIN1 has been linked to congenital generalized lipodystrophy and muscle-related phenotypes, making it a useful locus for studying membrane microdomain biology and metabolic regulation.
PTRF Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the CAVIN1 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within CAVIN1. 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 CAVIN1 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 CAVIN1-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.