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.
Chitotriosidase Double Nickase Plasmid (h): sc-402112-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Chitotriosidase 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
- Chitotriosidase Double Nickase Plasmid (h) and Chitotriosidase Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting CHIT1. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: Chitotriosidase Antibody (A-11): sc-271460
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Chitotriosidase Double Nickase Plasmid (h) | sc-402112-NIC | 20 µg | $410.00 | |||
Chitotriosidase Double Nickase Plasmid (h2) | sc-402112-NIC-2 | 20 µg | $410.00 |
CHIT1 encodes human chitotriosidase, a secreted glycosyl hydrolase produced primarily by activated macrophages and other myeloid cells that degrades chitin-like polysaccharides and participates in innate immune defense. CHIT1 is linked to lysosomal and phagolysosomal biology, macrophage activation programs, and extracellular matrix–associated remodeling processes that accompany chronic inflammation. Altered CHIT1 expression or activity is frequently used as a molecular readout of macrophage burden and inflammatory states, and genetic variation in CHIT1 can influence enzyme function in human populations. As a result, CHIT1 is relevant to studies of granulomatous and lysosomal storage disorders, airway inflammation and remodeling, and broader immunometabolic regulation of tissue homeostasis.
Chitotriosidase Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the CHIT1 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within CHIT1. 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 CHIT1 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 CHIT1-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.