![Su[fu] Antibody (F-4) - Western Blotting - Image 55082The Double Nickase Plasmid features a U6 promoter for sgRNA expression, a 20 nt targeting sequence, and a gRNA scaffold to guide Cas9n. It includes a CBh promoter for Cas9n (D10A) and puromycin resistance, GFP for transfection verification, and nuclear localization signals (NLS). The 2A peptide allows co-expression of Cas9n and Puro from a single promoter, enabling precise genome editing with reduced off-target effects.](https://media.scbt.com/product/sufu-double-nickase-plasmids-m-western-blotting_05_50_b_55082.jpg)
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.
Su[fu] Double Nickase Plasmid (m): sc-423973-NIC
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Su[fu] 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
- Su[fu] Double Nickase Plasmid (m) and Su[fu] Double Nickase Plasmid (m2) encode distinct paired gRNA designs targeting Sufu. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: Su[fu] Antibody (F-4): sc-137014
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Su[fu] Double Nickase Plasmid (m) | sc-423973-NIC | 20 µg | $410.00 | |||
Su[fu] Double Nickase Plasmid (m2) | sc-423973-NIC-2 | 20 µg | $410.00 |
Sufu (Suppressor of Fused) encodes a key intracellular inhibitor of Hedgehog signaling that restrains GLI transcription factors and helps set thresholds for pathway activation during embryonic patterning and tissue homeostasis. In mouse cells, Su[fu] regulates GLI processing, nuclear localization, and transcriptional output downstream of PTCH1/SMO, linking it to primary cilium–dependent signaling and developmental gene programs. Disruption of SUFU function is associated with aberrant Hedgehog pathway activity implicated in congenital malformations and tumorigenic processes, making Sufu a widely used node for studying signal transduction control. Sufu-centered models also support investigation of pathway cross-talk with cell cycle regulation, differentiation, and stem/progenitor maintenance.
Su[fu] Double Nickase Plasmid (m) consists of a matched pair of plasmids engineered for high-specificity editing of the Sufu locus in mouse cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within Sufu. 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 Sufu 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 Sufu-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.