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.
EWS Double Nickase Plasmid (h): sc-401036-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- EWS 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
- EWS Double Nickase Plasmid (h) and EWS Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting EWSR1. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: EWS Antibody (G-5): sc-28327
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
EWS Double Nickase Plasmid (h) | sc-401036-NIC | 20 µg | $410.00 | |||
EWS Double Nickase Plasmid (h2) | sc-401036-NIC-2 | 20 µg | $410.00 |
EWSR1 encodes the EWS RNA-binding protein, a multifunctional regulator of transcription and RNA processing that couples co-transcriptional splicing with RNA transport and stability. EWS participates in chromatin-associated transcriptional control and is linked to DNA damage responses and genome integrity through interactions with RNA polymerase II and RNA-processing complexes. Dysregulation of EWSR1, including chromosomal rearrangements that generate oncogenic fusion proteins, is a defining molecular feature in several sarcomas and other malignancies. Altered EWS function can perturb gene expression programs, RNA metabolism, and signaling networks that control proliferation and differentiation, making it a key target for mechanistic studies.
EWS Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the EWSR1 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within EWSR1. 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 EWSR1 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 EWSR1-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.