BLM hydrolase Double Nickase Plasmid (h): sc-404411-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- BLM hydrolase 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
- BLM hydrolase Double Nickase Plasmid (h) and BLM hydrolase Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting BLMH. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: BLM hydrolase Antibody (F-9): sc-166777
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
BLM hydrolase Double Nickase Plasmid (h) | sc-404411-NIC | 20 µg | $410.00 | |||
BLM hydrolase Double Nickase Plasmid (h2) | sc-404411-NIC-2 | 20 µg | $410.00 |
BLMH encodes bleomycin hydrolase, a cytosolic cysteine aminopeptidase that catalyzes N-terminal peptide processing and contributes to intracellular protein and peptide turnover. BLMH participates in general proteostasis and amino acid metabolism, and has been linked to processing of bioactive peptides, including amyloid-β, connecting its activity to pathways relevant to neurodegeneration and cellular stress responses. Altered BLMH expression or activity has been investigated in contexts of oxidative stress, inflammatory signaling, and differential sensitivity to bleomycin exposure, making it a useful target for mechanistic studies of detoxification and peptide catabolism. As a broadly expressed enzyme, BLMH serves as a model for dissecting how cytosolic peptidase function interfaces with protein quality control networks.
BLM hydrolase Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the BLMH locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within BLMH. 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 BLMH 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 BLMH-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.