LCAT Double Nickase Plasmid (h): sc-405225-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- LCAT 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
- LCAT Double Nickase Plasmid (h) and LCAT Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting LCAT. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: LCAT Antibody (D-2): sc-376682
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
LCAT Double Nickase Plasmid (h) | sc-405225-NIC | 20 µg | $410.00 | |||
LCAT Double Nickase Plasmid (h2) | sc-405225-NIC-2 | 20 µg | $410.00 |
Human LCAT (lecithin–cholesterol acyltransferase) is a secreted enzyme that catalyzes the esterification of free cholesterol on lipoprotein surfaces, generating cholesteryl esters that support HDL maturation and reverse cholesterol transport. By remodeling phospholipids and regulating lipoprotein particle composition, LCAT influences lipid homeostasis pathways linked to cholesterol efflux, apolipoprotein function, and systemic lipid trafficking. Altered LCAT activity is associated with dyslipidemia phenotypes and abnormal lipoprotein profiles, making it relevant for mechanistic studies of cardiometabolic risk biology. In experimental systems, LCAT perturbation is used to dissect relationships between HDL biogenesis, macrophage cholesterol handling, and lipid-driven inflammatory signaling.
LCAT Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the LCAT locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within LCAT. 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 LCAT 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 LCAT-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.