Ub-RPS27A Double Nickase Plasmid (h): sc-416542-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Ub-RPS27A 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
- Ub-RPS27A Double Nickase Plasmid (h) and Ub-RPS27A Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting RPS27A. One or both designs may be available
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Ub-RPS27A Double Nickase Plasmid (h) | sc-416542-NIC | 20 µg | $410.00 |
RPS27A encodes a fusion protein that is post-translationally processed to generate ubiquitin and the 40S ribosomal protein S27a, linking ubiquitin-dependent proteostasis with ribosome biogenesis and translation. Ubiquitin produced from Ub-RPS27A contributes to protein turnover via the ubiquitin–proteasome system, regulation of DNA damage signaling, and stress-responsive remodeling of the proteome. The S27a component is incorporated into the small ribosomal subunit and supports mRNA decoding and global translational capacity. Dysregulation of ribosomal proteins and ubiquitin homeostasis is implicated in altered cell growth control, proteotoxic stress responses, and genome maintenance pathways relevant to cancer and neurodegeneration research.
Ub-RPS27A Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the RPS27A locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within RPS27A. 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 RPS27A 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 RPS27A-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.