HuB Double Nickase Plasmid (h): sc-400169-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- HuB 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
- HuB Double Nickase Plasmid (h) and HuB Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting ELAVL2. One or both designs may be available
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
HuB Double Nickase Plasmid (h) | sc-400169-NIC | 20 µg | $410.00 | |||
HuB Double Nickase Plasmid (h2) | sc-400169-NIC-2 | 20 µg | $410.00 |
ELAVL2 (HuB) is a neuron-enriched RNA-binding protein of the Hu/ELAV family that recognizes AU-rich elements in target transcripts to regulate mRNA stability, alternative splicing, and translation. By shaping post-transcriptional gene expression programs, HuB influences neuronal differentiation, axon guidance, synaptic function, and activity-dependent plasticity, intersecting with pathways controlling RNA processing and local translation. Dysregulated ELAVL2 expression or function has been linked to altered neuronal gene networks observed in neurodevelopmental and neurodegenerative contexts, making it relevant for studying mechanisms that couple RNA metabolism to neuronal phenotype. Experimental modulation of HuB provides a handle to interrogate how RNA-binding proteins coordinate transcriptome dynamics in human neural models.
HuB Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the ELAVL2 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within ELAVL2. 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 ELAVL2 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 ELAVL2-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.