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.
CIN85 Double Nickase Plasmid (h): sc-402573-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- CIN85 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
- CIN85 Double Nickase Plasmid (h) and CIN85 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting SH3KBP1. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: CIN85 Antibody (A-7): sc-166862
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
CIN85 Double Nickase Plasmid (h) | sc-402573-NIC | 20 µg | $410.00 |
SH3KBP1 encodes CIN85, a multi-adaptor scaffold protein that coordinates endocytic trafficking and signal attenuation by assembling complexes downstream of receptor tyrosine kinases, including EGFR, via interactions with Cbl ubiquitin ligases and endocytic machinery. Through its SH3 domains and proline-rich binding interfaces, CIN85 regulates receptor internalization, ubiquitination, and lysosomal sorting, shaping the amplitude and duration of MAPK and PI3K-AKT signaling. CIN85 also contributes to cytoskeletal remodeling, invadopodia formation, and vesicle dynamics, processes linked to cellular migration and invasive phenotypes in multiple disease contexts. Dysregulated CIN85-associated pathways have been studied in relation to oncogenic signaling, immune receptor turnover, and altered membrane trafficking.
CIN85 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the SH3KBP1 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within SH3KBP1. 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 SH3KBP1 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 SH3KBP1-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.