Alox12e CRISPR/Cas9 KO Plasmid (m): sc-419087
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Alox12e CRISPR/Cas9 Knockout (KO) Plasmid (m) 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 Alox12e 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
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Alox12e CRISPR/Cas9 KO Plasmid (m) | sc-419087 | 20 µg | $397.00 |
Overview
Alox12e encodes arachidonate 12-lipoxygenase, 12R-type, a non-heme iron dioxygenase that catalyzes oxygenation of polyunsaturated fatty acids to generate bioactive lipid mediators. In mouse epidermis, Alox12e functions within the lipoxygenase pathway and cooperates with related enzymes in oxidative lipid processing that supports keratinocyte differentiation and formation of the stratum corneum barrier. Through regulation of lipid oxidation products and downstream signaling, Alox12e contributes to barrier homeostasis, inflammatory responses, and stress signaling in skin. Disruption of this axis is relevant to studies of ichthyosis-like barrier defects, dermatitis-associated inflammation, and broader lipid metabolism phenotypes in epithelial biology.
Alox12e CRISPR/Cas9 KO Plasmid (m) is a pool of plasmids designed for targeted disruption of the Alox12e gene in mouse cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the Alox12e 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 Alox12e 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 Alox12e protein expression.
This CRISPR knockout system enables efficient generation of Alox12e-deficient cell models for investigation of Alox12e signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting Alox12e exon(s) critical for Alox12e 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 Alox12e genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by Alox12e CRISPR/Cas9 KO Plasmid (m) and Alox12e CRISPR/Cas9 KO Plasmid (m2) target distinct sites within the Alox12e locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by Alox12e HDR Plasmid (m) and Alox12e HDR Plasmid (m2) contain a puromycin resistance cassette and an RFP reporter flanked by Alox12e homology arms to support homology-directed repair at defined Alox12e 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.