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.
Rad54 Double Nickase Plasmid (h): sc-401750-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Rad54 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
- Rad54 Double Nickase Plasmid (h) and Rad54 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting RAD54L. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: Rad54 Antibody (F-11): sc-374598
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Rad54 Double Nickase Plasmid (h) | sc-401750-NIC | 20 µg | $410.00 | |||
Rad54 Double Nickase Plasmid (h2) | sc-401750-NIC-2 | 20 µg | $410.00 |
RAD54L encodes the human Rad54 DNA-dependent ATPase, a core component of homologous recombination that remodels chromatin and promotes RAD51-mediated strand invasion during repair of DNA double-strand breaks. Rad54 functions within the Fanconi anemia/BRCA DNA repair network to support replication fork stability, facilitate post-replication repair, and preserve genome integrity under genotoxic stress. Disruption of RAD54L perturbs high-fidelity recombination and can shift repair toward error-prone pathways, contributing to chromosomal rearrangements and mutational accumulation observed in cancer biology. RAD54L is therefore widely used as a mechanistic entry point for studying DNA damage response signaling, recombination dynamics, and determinants of genomic instability.
Rad54 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the RAD54L locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within RAD54L. 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 RAD54L 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 RAD54L-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.