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.
RPA194 Double Nickase Plasmid (h): sc-400816-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- RPA194 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
- RPA194 Double Nickase Plasmid (h) and RPA194 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting POLR1A. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: RPA194 Antibody (C-1): sc-48385
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
RPA194 Double Nickase Plasmid (h) | sc-400816-NIC | 20 µg | $410.00 | |||
RPA194 Double Nickase Plasmid (h2) | sc-400816-NIC-2 | 20 µg | $410.00 |
POLR1A encodes RPA194, the catalytic subunit of RNA polymerase I that drives 47S pre-rRNA synthesis in the nucleolus, a rate-limiting step for ribosome biogenesis. RPA194 functions within the Pol I transcription machinery to support nucleolar organization, cell growth, and coordination of ribosomal RNA processing with cellular metabolic state. Perturbation of POLR1A-dependent rRNA transcription alters nucleolar stress signaling and downstream p53-associated responses, linking this pathway to cell-cycle control and genome surveillance. Genetic variation and dysregulation of Pol I activity have been associated with ribosomopathy-related phenotypes and are widely studied in the context of proliferative biology and stress adaptation.
RPA194 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the POLR1A locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within POLR1A. 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 POLR1A 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 POLR1A-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.