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.
TFIIH p80 Double Nickase Plasmid (h): sc-402231-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- TFIIH p80 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 p80 Double Nickase Plasmid (h) and TFIIH p80 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting ERCC2. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: TFIIH p80 Antibody (G-2): sc-271206
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
TFIIH p80 Double Nickase Plasmid (h) | sc-402231-NIC | 20 µg | $410.00 | |||
TFIIH p80 Double Nickase Plasmid (h2) | sc-402231-NIC-2 | 20 µg | $410.00 |
ERCC2 encodes the human TFIIH p80 helicase (XPD), an ATP-dependent 5′–3′ DNA helicase that is an essential subunit of the TFIIH complex. TFIIH coordinates nucleotide excision repair by unwinding damaged DNA around bulky lesions and also supports RNA polymerase II transcription initiation through promoter opening. ERCC2 activity links genome stability pathways with transcription-coupled repair, and defects in TFIIH function are associated with UV sensitivity and syndromic DNA repair disorders including xeroderma pigmentosum, Cockayne syndrome, and trichothiodystrophy.
TFIIH p80 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the ERCC2 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within ERCC2. 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 ERCC2 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 ERCC2-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.