ITM2B Double Nickase Plasmid (h): sc-403561-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- ITM2B 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
- ITM2B Double Nickase Plasmid (h) and ITM2B Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting ITM2B. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: ITM2B Antibody (C-8): sc-374362
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
ITM2B Double Nickase Plasmid (h) | sc-403561-NIC | 20 µg | $410.00 | |||
ITM2B Double Nickase Plasmid (h2) | sc-403561-NIC-2 | 20 µg | $410.00 |
ITM2B encodes an integral membrane protein of the BRICHOS domain family that traffics through the secretory and endolysosomal systems and is enriched in neurons. ITM2B contributes to membrane protein processing and proteostasis, with reported roles in regulating amyloid precursor protein processing, vesicular transport, and neurite-associated cellular homeostasis. Disruption of ITM2B function is linked to neurodegenerative phenotypes, including familial British and familial Danish dementias, and it has been studied in the context of amyloidogenic peptide generation and intracellular stress responses. These properties make ITM2B a relevant target for investigating pathways connecting membrane trafficking, protein quality control, and neuronal vulnerability.
ITM2B Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the ITM2B locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within ITM2B. 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 ITM2B 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 ITM2B-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.