TFIIH p89 Double Nickase Plasmid (h): sc-401124-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- TFIIH p89 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
- TFIIH p89 Double Nickase Plasmid (h) and TFIIH p89 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting ERCC3. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: TFIIH p89 Antibody (G-10): sc-271500
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
TFIIH p89 Double Nickase Plasmid (h) | sc-401124-NIC | 20 µg | $410.00 | |||
TFIIH p89 Double Nickase Plasmid (h2) | sc-401124-NIC-2 | 20 µg | $410.00 |
ERCC3 encodes TFIIH p89 (XPB), a DNA-dependent ATPase and 3′→5′ helicase core to the TFIIH complex that couples basal transcription initiation by RNA polymerase II with nucleotide excision repair. By unwinding promoter DNA and lesion-containing DNA during transcription-coupled and global genome NER, TFIIH p89 helps coordinate genome integrity, cell-cycle progression, and responses to UV-induced damage. Disruption of ERCC3 function perturbs transcriptional homeostasis and DNA repair capacity, linking ERCC3 defects to inherited DNA repair syndromes such as xeroderma pigmentosum and trichothiodystrophy. As a TFIIH component, TFIIH p89 is frequently studied in mechanisms of transcription regulation, replication stress, and mutagenesis pathways.
TFIIH p89 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the ERCC3 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within ERCC3. 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 ERCC3 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 ERCC3-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.