Epidermis-type 12-LO CRISPR/Cas9 KO Plasmid (m): sc-419088
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Epidermis-type 12-LO 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 Epidermis-type 12-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
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Epidermis-type 12-LO CRISPR/Cas9 KO Plasmid (m) | sc-419088 | 20 µg | $397.00 |
Overview
Alox12b encodes epidermis-type 12-lipoxygenase (12-LO), a non-heme iron-dependent dioxygenase that oxidizes esterified polyunsaturated fatty acids to generate hydroperoxy lipid intermediates. In mouse epidermis, ALOX12B functions within the lipoxygenase pathway that supports acylceramide processing and formation of the stratum corneum lipid barrier, coordinating terminal keratinocyte differentiation and cornification. Altered 12-LO activity impacts epidermal lipid homeostasis, oxidative lipid signaling, and barrier integrity, processes commonly studied in models of ichthyosis-like phenotypes, dermatitis, and inflammatory skin stress. As a result, Alox12b is a useful entry point for dissecting lipid-metabolic contributions to epithelial barrier function and cutaneous immune signaling.
Epidermis-type 12-LO CRISPR/Cas9 KO Plasmid (m) is a pool of plasmids designed for targeted disruption of the Alox12b gene in mouse cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the Alox12b 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 Alox12b 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 Epidermis-type 12-LO protein expression.
This CRISPR knockout system enables efficient generation of Alox12b-deficient cell models for investigation of Epidermis-type 12-LO signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting Alox12b exon(s) critical for Epidermis-type 12-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 Alox12b genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by Epidermis-type 12-LO CRISPR/Cas9 KO Plasmid (m) and Epidermis-type 12-LO CRISPR/Cas9 KO Plasmid (m2) target distinct sites within the Alox12b locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by Epidermis-type 12-LO HDR Plasmid (m) and Epidermis-type 12-LO HDR Plasmid (m2) contain a puromycin resistance cassette and an RFP reporter flanked by Alox12b homology arms to support homology-directed repair at defined Alox12b 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.