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.
5-LO CRISPR/Cas9 KO Plasmid (h): sc-401239
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- 5-LO 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 5-LO 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: 5-LO Antibody (33): sc-136195
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
5-LO CRISPR/Cas9 KO Plasmid (h) | sc-401239 | 20 µg | $397.00 |
Overview
ALOX5 encodes 5-lipoxygenase (5-LO), a non-heme iron dioxygenase that catalyzes the first committed steps in arachidonic acid metabolism to leukotriene A4, which is subsequently converted into bioactive leukotrienes. 5-LO activity is regulated by cellular calcium flux, membrane association with FLAP (ALOX5AP), phosphorylation-dependent signaling, and redox status, linking it to innate immune activation and inflammatory lipid mediator networks. Leukotriene signaling through CysLT and BLT receptors shapes leukocyte chemotaxis, vascular permeability, and cytokine release, positioning ALOX5 within pathways relevant to asthma and allergic inflammation, atherosclerosis, and tumor-associated inflammation. Dysregulated ALOX5 expression or pathway flux is also used as a mechanistic readout in studies of oxidative stress, macrophage polarization, and eicosanoid crosstalk with cyclooxygenase-derived prostanoids.
5-LO CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the ALOX5 gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the ALOX5 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 ALOX5 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 5-LO protein expression.
This CRISPR knockout system enables efficient generation of ALOX5-deficient cell models for investigation of 5-LO signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting ALOX5 exon(s) critical for 5-LO 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 ALOX5 genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by 5-LO CRISPR/Cas9 KO Plasmid (h) and 5-LO CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the ALOX5 locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by 5-LO HDR Plasmid (h) and 5-LO HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by ALOX5 homology arms to support homology-directed repair at defined ALOX5 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.