HECTD1 Double Nickase Plasmid (h): sc-409864-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- HECTD1 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
- HECTD1 Double Nickase Plasmid (h) and HECTD1 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting HECTD1. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: HECTD1 Antibody (1E10): sc-517169
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
HECTD1 Double Nickase Plasmid (h) | sc-409864-NIC | 20 µg | $410.00 | |||
HECTD1 Double Nickase Plasmid (h2) | sc-409864-NIC-2 | 20 µg | $410.00 |
HECTD1 encodes a HECT-domain E3 ubiquitin ligase that regulates protein ubiquitination and proteasome-dependent turnover, shaping signal transduction and cytoskeletal organization. As a modulator of ubiquitin-driven quality control, HECTD1 has been linked to processes including stress-response signaling, cell migration, and developmental regulation through control of substrate stability and trafficking. Perturbation of HECTD1 activity can alter pathway dynamics that influence cell-cycle progression and morphogenetic programs, making it relevant to studies of dysregulated growth and tissue patterning. Its ubiquitin ligase function also positions HECTD1 as a useful node for investigating crosstalk between ubiquitination, protein homeostasis, and transcriptional responses.
HECTD1 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the HECTD1 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within HECTD1. 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 HECTD1 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 HECTD1-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.