CIS Double Nickase Plasmid (h): sc-402619-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- CIS 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
- CIS Double Nickase Plasmid (h) and CIS Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting CISH. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: CIS Antibody (D-8): sc-74581
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
CIS Double Nickase Plasmid (h) | sc-402619-NIC | 20 µg | $410.00 | |||
CIS Double Nickase Plasmid (h2) | sc-402619-NIC-2 | 20 µg | $410.00 |
CISH encodes cytokine-inducible SH2-containing protein (CIS), a member of the SOCS family that provides negative feedback control of cytokine receptor signaling. CIS binds phosphorylated receptor complexes and can promote ubiquitin-mediated turnover, thereby limiting JAK/STAT pathway amplitude and duration in immune and hematopoietic cells. Through this regulatory role, CIS influences T cell activation, NK cell responsiveness, and cytokine-driven differentiation programs. Altered CISH expression or regulation has been associated with immune dysregulation and cancer-related signaling contexts, making it a useful node for dissecting cytokine signaling networks.
CIS Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the CISH locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within CISH. 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 CISH 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 CISH-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.