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.
ISG15 Double Nickase Plasmid (h): sc-400996-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- ISG15 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
- ISG15 Double Nickase Plasmid (h) and ISG15 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting ISG15. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: ISG15 Antibody (F-9): sc-166755
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
ISG15 Double Nickase Plasmid (h) | sc-400996-NIC | 20 µg | $410.00 | |||
ISG15 Double Nickase Plasmid (h2) | sc-400996-NIC-2 | 20 µg | $410.00 |
ISG15 encodes an interferon-stimulated ubiquitin-like modifier that is rapidly induced by type I interferons and cellular stress. Conjugation of ISG15 to protein substrates (ISGylation) is mediated by the E1 enzyme UBA7, E2 UBE2L6, and E3 ligases such as HERC5, and is reversed by the deISGylase USP18, integrating ISG15 into innate immune and proteostasis networks. ISG15 impacts antiviral restriction, regulation of interferon signaling amplitude, and modulation of translation and protein turnover, with additional extracellular cytokine-like activities reported in some contexts. Dysregulated ISG15/ISGylation has been associated with altered host–pathogen interactions, interferonopathies, inflammatory phenotypes, and cancer-related immune signaling, making it a relevant target for mechanistic studies of interferon pathways.
ISG15 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the ISG15 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within ISG15. 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 ISG15 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 ISG15-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.