TFIIIB90-1 Double Nickase Plasmid (h): sc-405826-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- TFIIIB90-1 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
- TFIIIB90-1 Double Nickase Plasmid (h) and TFIIIB90-1 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting BRF1. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: TFIIIB90-1/2/3/5 Antibody (A-8): sc-390821
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
TFIIIB90-1 Double Nickase Plasmid (h) | sc-405826-NIC | 20 µg | $410.00 |
BRF1 encodes TFIIIB90-1, a core subunit of the RNA polymerase III transcription factor TFIIIB that cooperates with TBP and BDP1 to position Pol III at promoters of tRNA genes, 5S rRNA, and other small noncoding RNA loci. By controlling Pol III initiation, TFIIIB90-1 influences ribosome biogenesis, translational capacity, and cellular growth programs, linking nutrient sensing and stress-responsive transcriptional regulation to proliferation. Dysregulated Pol III output and TFIIIB component activity have been associated with oncogenic transformation and altered proteostasis in multiple contexts, making BRF1 a useful node for dissecting transcriptional control of biosynthetic pathways. Functional studies of BRF1 commonly intersect with chromatin regulation at Pol III promoters, cell-cycle progression, and metabolic signaling that modulates Pol III transcriptional throughput.
TFIIIB90-1 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the BRF1 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within BRF1. 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 BRF1 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 BRF1-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.