NRSF Double Nickase Plasmid (h): sc-418578-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- NRSF 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
- NRSF Double Nickase Plasmid (h) and NRSF Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting REST. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: NRSF Antibody (F-3): sc-374611
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
NRSF Double Nickase Plasmid (h) | sc-418578-NIC | 20 µg | $410.00 | |||
NRSF Double Nickase Plasmid (h2) | sc-418578-NIC-2 | 20 µg | $410.00 |
REST, also known as neuron-restrictive silencer factor (NRSF), is a zinc-finger transcriptional repressor that binds repressor element-1 (RE1/NRSE) sites to silence neuronal gene programs in non-neuronal cells and modulate neuronal maturation. Through recruitment of corepressor complexes such as SIN3/HDAC and CoREST/LSD1, REST reshapes chromatin to regulate synaptic genes, ion channels, and neurosecretory pathways, influencing differentiation, excitability, and stress responses. REST-dependent transcriptional control intersects with epigenetic regulation and activity-dependent signaling, contributing to context-specific shifts in cell identity and plasticity. Dysregulated REST/NRSF activity has been associated with neurodevelopmental and neurodegenerative mechanisms as well as aberrant transcriptional repression programs observed in multiple tumor contexts, supporting its use as a node for studying gene regulatory networks.
NRSF Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the REST locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within REST. 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 REST 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 REST-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.