Rad18 Double Nickase Plasmid (h): sc-406099-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Rad18 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
- Rad18 Double Nickase Plasmid (h) and Rad18 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting RAD18. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: Rad18 Antibody (79B1048): sc-52949
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Rad18 Double Nickase Plasmid (h) | sc-406099-NIC | 20 µg | $410.00 |
Human RAD18 encodes Rad18, an E3 ubiquitin ligase that coordinates post-replication DNA damage tolerance by partnering with the E2 enzyme RAD6 to monoubiquitinate PCNA and promote translesion synthesis and template switching. Through this PCNA-centered pathway, Rad18 helps restart stalled replication forks and limits replication-associated genome instability following UV and other genotoxic stresses. RAD18 interfaces with broader DNA repair and replication stress response networks, influencing mutagenesis rates, chromosomal stability, and cellular sensitivity to DNA damaging agents. Dysregulation of RAD18 has been associated with altered DNA damage tolerance capacity and genomic instability phenotypes that are frequently studied in cancer biology and inherited DNA repair disorder models.
Rad18 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the RAD18 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within RAD18. 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 RAD18 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 RAD18-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.