TFIIB Double Nickase Plasmid (h): sc-401174-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- TFIIB 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
- TFIIB Double Nickase Plasmid (h) and TFIIB Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting GTF2B. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: TFIIB Antibody (D-3): sc-271736
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
TFIIB Double Nickase Plasmid (h) | sc-401174-NIC | 20 µg | $410.00 | |||
TFIIB Double Nickase Plasmid (h2) | sc-401174-NIC-2 | 20 µg | $410.00 |
Human GTF2B encodes transcription factor IIB (TFIIB), a core component of the RNA polymerase II preinitiation complex that bridges TBP/TFIID at the TATA box with Pol II and helps position the transcription start site. TFIIB supports promoter recognition, transcription initiation, and early promoter escape, influencing global mRNA output and cellular state transitions. Through its central role in Pol II-dependent transcription, perturbation of TFIIB function can impact pathways controlling proliferation, differentiation, and stress responses. Dysregulation of the general transcription machinery, including factors that modulate TFIIB-dependent initiation dynamics, has been implicated in oncogenic transcription programs and other diseases driven by altered gene expression.
TFIIB Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the GTF2B locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within GTF2B. 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 GTF2B 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 GTF2B-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.