Not CRISPR data: the image illustrates antibody quality for visualizing CRISPR experiment results.
Not CRISPR data: the image illustrates antibody quality for visualizing CRISPR experiment results.
RBCK1 Double Nickase Plasmid (h): sc-402044-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- RBCK1 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
- RBCK1 Double Nickase Plasmid (h) and RBCK1 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting RBCK1. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: RBCK1 Antibody (E-2): sc-365523
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
RBCK1 Double Nickase Plasmid (h) | sc-402044-NIC | 20 µg | $410.00 | |||
RBCK1 Double Nickase Plasmid (h2) | sc-402044-NIC-2 | 20 µg | $410.00 |
RBCK1 (also known as HOIL-1) encodes an E3 ubiquitin ligase that functions as a core component of the linear ubiquitin chain assembly complex (LUBAC), coordinating Met1-linked ubiquitination events that shape innate and adaptive immune signaling. Through regulation of NF-κB activation and inflammatory pathway outputs downstream of receptors such as TNFR and certain pattern-recognition receptors, RBCK1 influences cytokine production, cell survival, and stress responses. RBCK1 also participates in proteostasis and quality-control pathways via ubiquitin signaling networks. Dysregulation of RBCK1/LUBAC activity has been linked to immune and autoinflammatory phenotypes and has been studied in contexts including immunodeficiency and tissue inflammation.
RBCK1 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the RBCK1 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within RBCK1. 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 RBCK1 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 RBCK1-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.