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.
Hrs Double Nickase Plasmid (h): sc-401501-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Hrs 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
- Hrs Double Nickase Plasmid (h) and Hrs Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting HGS. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: Hrs Antibody (C-7): sc-271455
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Hrs Double Nickase Plasmid (h) | sc-401501-NIC | 20 µg | $410.00 | |||
Hrs Double Nickase Plasmid (h2) | sc-401501-NIC-2 | 20 µg | $410.00 |
Human HGS encodes hepatocyte growth factor–regulated tyrosine kinase substrate (Hrs), an ESCRT-0 component that initiates endosomal sorting of ubiquitinated membrane proteins. Hrs binds phosphatidylinositol 3-phosphate on early endosomes and coordinates cargo recognition with ESCRT-I/II/III assemblies to drive multivesicular body formation, lysosomal degradation, and down-modulation of receptor signaling. Through these activities, Hrs influences endocytosis-dependent control of pathways such as EGFR and other receptor tyrosine kinase networks, impacting signal duration and cellular homeostasis. Dysregulated endosomal trafficking and receptor turnover associated with altered HGS/Hrs function has been linked to contexts relevant to cancer biology and neurodegenerative disease mechanisms, making it a useful node for studying proteostasis and signaling adaptation.
Hrs Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the HGS locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within HGS. 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 HGS 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 HGS-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.