Lyst Double Nickase Plasmid (m): sc-421511-NIC
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Lyst Double Nickase Plasmid (m) 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
- Lyst Double Nickase Plasmid (m) and Lyst Double Nickase Plasmid (m2) encode distinct paired gRNA designs targeting Lyst. One or both designs may be available
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Lyst Double Nickase Plasmid (m) | sc-421511-NIC | 20 µg | $410.00 |
Mouse Lyst (lysosomal trafficking regulator) encodes a large cytosolic protein essential for lysosome-related organelle biogenesis and vesicle fusion–fission dynamics. It coordinates endolysosomal trafficking, regulates lysosome size and distribution, and supports proper maturation of phagosomes and secretory granules in immune and pigment cells. Disruption of Lyst perturbs autophagy-lysosome function and antigen processing, leading to defects in degranulation and intracellular killing. LYST dysfunction is linked to Chediak–Higashi syndrome–like phenotypes, making Lyst a key gene for studying immune dysregulation, pigmentation biology, and lysosome-associated pathways.
Lyst Double Nickase Plasmid (m) consists of a matched pair of plasmids engineered for high-specificity editing of the Lyst locus in mouse cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within Lyst. 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 Lyst 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 Lyst-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.