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.
NTE CRISPR/Cas9 KO Plasmid (h): sc-406850
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- NTE CRISPR/Cas9 Knockout (KO) Plasmid (h) is a pool of plasmids, each encoding Cas9 nuclease and a target-specific 20 nt guide RNA (gRNA) designed for maximum knockout efficiency using sequences derived from the GeCKO v2 library
- gRNA sequences direct Cas9 to induce site-specific double-strand breaks (DSBs) in the NTE genomic locus, resulting in gene knockout through non-homologous end joining (NHEJ)
- The puromycin resistance and RFP genes are flanked by LoxP sites, enabling removal of selection markers via Cre recombinase (Cre Vector: sc-418923) after establishing stable knockout cell lines
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: NTE Antibody (G-4): sc-271049
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
NTE CRISPR/Cas9 KO Plasmid (h) | sc-406850 | 20 µg | $397.00 |
Overview
PNPLA6 encodes neuropathy target esterase (NTE), a membrane-associated phospholipase/lysophospholipase that hydrolyzes phosphatidylcholine to regulate phospholipid homeostasis. NTE activity supports endoplasmic reticulum membrane integrity, lipid droplet dynamics, and neuronal maintenance by controlling levels of bioactive lysophospholipids. Disruption of PNPLA6 perturbs membrane remodeling and lipid signaling pathways, processes closely linked to axon stability and synaptic function. Genetic variation in PNPLA6 is associated with neurodegenerative and neurodevelopmental phenotypes, making it a useful locus for studying lipid-driven mechanisms of neuronal vulnerability.
NTE CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the PNPLA6 gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the PNPLA6 together with the Streptococcus pyogenes Cas9 nuclease. The plasmids also encode GFP, allowing fluorescent identification and enrichment of successfully transfected cells by fluorescence microscopy or flow cytometry.
The multi-guide design increases the likelihood of generating insertions or deletions (indels) that disrupt the PNPLA6 open reading frame following Cas9-mediated double-strand break formation. DNA breaks introduced by the CRISPR/Cas9 system are repaired through endogenous non-homologous end joining (NHEJ) pathways, frequently resulting in frameshift mutations that abolish NTE protein expression.
This CRISPR knockout system enables efficient generation of PNPLA6-deficient cell models for investigation of NTE signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting PNPLA6 exon(s) critical for NTE function
- Co-expression of SpCas9 and sgRNA from a single plasmid for simplified delivery
- GFP reporter for identification of transfected cells
- Pool of plasmids targeting multiple PNPLA6 genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by NTE CRISPR/Cas9 KO Plasmid (h) and NTE CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the PNPLA6 locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by NTE HDR Plasmid (h) and NTE HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by PNPLA6 homology arms to support homology-directed repair at defined PNPLA6 target sites corresponding to the CRISPR/Cas9 KO designs. HDR donor availability may vary. See Related Products for availability.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.