RBM35A Double Nickase Plasmid (h): sc-405645-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- RBM35A 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
- RBM35A Double Nickase Plasmid (h) and RBM35A Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting ESRP1. One or both designs may be available
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
RBM35A Double Nickase Plasmid (h) | sc-405645-NIC | 20 µg | $410.00 | |||
RBM35A Double Nickase Plasmid (h2) | sc-405645-NIC-2 | 20 µg | $410.00 |
ESRP1 (RBM35A) is an epithelial splicing regulatory protein that binds pre-mRNA and directs alternative splicing programs that distinguish epithelial from mesenchymal transcript isoforms. It helps maintain epithelial identity by controlling splice choices in genes involved in cell–cell adhesion, cytoskeletal organization, and signal transduction, influencing processes such as EMT and epithelial differentiation. Through coordinated regulation of exon inclusion and exclusion, ESRP1 shapes pathways linked to cell polarity and migration, and its dysregulation is frequently studied in the context of developmental transitions and tumor-associated phenotypic switching. Altered ESRP1-dependent splicing networks have been associated with changes in invasion-related isoforms and lineage plasticity, making it a common target in mechanistic studies of RNA processing and cancer biology.
RBM35A Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the ESRP1 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within ESRP1. 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 ESRP1 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 ESRP1-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.