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.
CLN1 Double Nickase Plasmid (h): sc-404018-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- CLN1 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
- CLN1 Double Nickase Plasmid (h) and CLN1 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting PPT1. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: CLN1 Antibody (1117CT11.2.1.4): sc-517323
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
CLN1 Double Nickase Plasmid (h) | sc-404018-NIC | 20 µg | $410.00 | |||
CLN1 Double Nickase Plasmid (h2) | sc-404018-NIC-2 | 20 µg | $410.00 |
PPT1 encodes palmitoyl-protein thioesterase 1 (CLN1), a lysosomal hydrolase that removes long-chain fatty acyl groups from S-palmitoylated proteins, enabling their turnover and recycling. By regulating depalmitoylation, CLN1 supports lysosome-dependent proteostasis, endolysosomal trafficking, and autophagy-lysosome pathway dynamics, with downstream effects on neuronal maintenance and synaptic function. Loss of PPT1 activity leads to accumulation of lipid-modified substrates and lysosomal storage material, linking this pathway to neuronal ceroid lipofuscinosis biology and broader mechanisms of neurodegeneration and cellular stress responses.
CLN1 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the PPT1 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within PPT1. 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 PPT1 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 PPT1-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.