TFEB Double Nickase Plasmid (m): sc-437332-NIC
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- TFEB 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
- TFEB Double Nickase Plasmid (m) and TFEB Double Nickase Plasmid (m2) encode distinct paired gRNA designs targeting Tfeb. One or both designs may be available
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
TFEB Double Nickase Plasmid (m) | sc-437332-NIC | 20 µg | $410.00 |
Mouse Tfeb encodes TFEB, a basic helix–loop–helix leucine zipper transcription factor that acts as a master regulator of lysosomal biogenesis and autophagy by coordinating the CLEAR gene network. TFEB activity is tightly controlled by nutrient- and stress-sensing pathways, including mTORC1-dependent phosphorylation that influences cytoplasmic sequestration versus nuclear translocation, thereby tuning lysosome function, autophagic flux, and cellular clearance. Through these programs, TFEB shapes proteostasis, mitochondrial quality control, and innate immune signaling, linking environmental cues to catabolic remodeling. Dysregulated TFEB signaling has been associated with lysosomal storage phenotypes, neurodegeneration-related proteotoxic stress, metabolic imbalance, and tumor cell adaptation to stress, making Tfeb a key target for mechanistic studies of cellular homeostasis.
TFEB Double Nickase Plasmid (m) consists of a matched pair of plasmids engineered for high-specificity editing of the Tfeb locus in mouse cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within Tfeb. 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 Tfeb 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 Tfeb-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.