Sacsin Double Nickase Plasmid (m): sc-424466-NIC
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Sacsin 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
- Sacsin Double Nickase Plasmid (m) and Sacsin Double Nickase Plasmid (m2) encode distinct paired gRNA designs targeting Sacs. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: Sacsin Antibody (G-3): sc-515118
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Sacsin Double Nickase Plasmid (m) | sc-424466-NIC | 20 µg | $410.00 |
Mouse Sacs encodes sacsin, a large cytosolic co-chaperone implicated in protein quality control and cytoskeletal homeostasis. Sacsin interfaces with Hsp70/Hsp90 chaperone networks and is linked to regulation of intermediate filaments, axonal transport, and mitochondrial dynamics, supporting neuronal proteostasis under stress. Disruption of sacsin function is associated with neurodegenerative phenotypes characterized by impaired motor coordination and progressive neuronal vulnerability, making Sacs a useful locus for modeling pathways of proteotoxic stress and axon maintenance.
Sacsin Double Nickase Plasmid (m) consists of a matched pair of plasmids engineered for high-specificity editing of the Sacs locus in mouse cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within Sacs. 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 Sacs 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 Sacs-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.