HuD Double Nickase Plasmid (h): sc-400132-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- HuD 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
- HuD Double Nickase Plasmid (h) and HuD Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting ELAVL4. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: HuD Antibody (H-9): sc-48421
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
HuD Double Nickase Plasmid (h) | sc-400132-NIC | 20 µg | $410.00 | |||
HuD Double Nickase Plasmid (h2) | sc-400132-NIC-2 | 20 µg | $410.00 |
ELAVL4 encodes HuD, a neuron-enriched RNA-binding protein of the ELAV/Hu family that recognizes AU-rich elements to regulate mRNA stability, alternative splicing, and translation. HuD coordinates post-transcriptional gene control programs that support neuronal differentiation, axonal growth, synaptic plasticity, and activity-dependent remodeling, integrating with stress granule dynamics and RNA transport in neurites. Dysregulated ELAVL4/HuD function and expression have been linked to altered neuronal maturation and RNA metabolism in neurodevelopmental and neurodegenerative contexts, and ELAVL4 is also used as a neuronal lineage marker in brain tumor biology and paraneoplastic neurological syndromes. These features make ELAVL4 a useful node for studying neuronal RNA regulons, transcript isoform selection, and protein–RNA network rewiring.
HuD Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the ELAVL4 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within ELAVL4. 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 ELAVL4 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 ELAVL4-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.