AMBRA1 Double Nickase Plasmid (h): sc-404108-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- AMBRA1 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
- AMBRA1 Double Nickase Plasmid (h) and AMBRA1 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting AMBRA1. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: AMBRA1 Antibody (G-6): sc-398204
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
AMBRA1 Double Nickase Plasmid (h) | sc-404108-NIC | 20 µg | $410.00 | |||
AMBRA1 Double Nickase Plasmid (h2) | sc-404108-NIC-2 | 20 µg | $410.00 |
AMBRA1 (autophagy and beclin 1 regulator 1) encodes a scaffold protein that coordinates autophagy initiation by modulating the BECLIN1/PIK3C3 complex and integrating signals from nutrient and stress pathways. AMBRA1 also interfaces with ubiquitin-dependent proteostasis and cell-cycle control, contributing to cellular homeostasis through regulation of mitochondrial quality control and apoptosis-related signaling. Dysregulation of AMBRA1-linked autophagy networks has been associated with altered neuronal development and tumor-relevant processes, making it a useful node for studying stress adaptation and proteostasis imbalance in human cells. As an autophagy regulator with broad pathway connectivity, AMBRA1 is frequently interrogated in models of neurobiology, metabolism, and cancer cell signaling.
AMBRA1 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the AMBRA1 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within AMBRA1. 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 AMBRA1 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 AMBRA1-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.