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.
CSA Double Nickase Plasmid (h): sc-403428-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- CSA 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
- CSA Double Nickase Plasmid (h) and CSA Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting ERCC8. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: CSA Antibody (D-2): sc-376981
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
CSA Double Nickase Plasmid (h) | sc-403428-NIC | 20 µg | $410.00 | |||
CSA Double Nickase Plasmid (h2) | sc-403428-NIC-2 | 20 µg | $410.00 |
ERCC8 encodes the human CSA protein, a WD40-repeat factor that forms the CRL4CSA E3 ubiquitin ligase complex with DDB1, CUL4A, and RBX1 to coordinate transcription-coupled nucleotide excision repair (TC-NER). CSA helps resolve RNA polymerase II–stalling lesions by recruiting repair and chromatin-remodeling activities and promoting ubiquitin-dependent turnover of damage-response components. Through these functions, CSA supports genome stability, recovery of transcription after UV-induced damage, and maintenance of cellular homeostasis under genotoxic stress. Pathogenic ERCC8 disruption is linked to Cockayne syndrome group A, a DNA repair disorder used as a model to study TC-NER defects, transcription stress, and neurodevelopmental vulnerability.
CSA Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the ERCC8 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within ERCC8. 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 ERCC8 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 ERCC8-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.