α-gal A CRISPR/Cas9 KO Plasmid (m2): sc-419046-KO-2
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- α-gal A CRISPR/Cas9 Knockout (KO) Plasmid (m2) 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 α-gal A 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 | |
α-gal A CRISPR/Cas9 KO Plasmid (m2) | sc-419046-KO-2 | 20 µg | $397.00 |
Overview
Gla encodes mouse α-galactosidase A (α-gal A), a lysosomal exoglycosidase that removes terminal α-galactosyl residues from glycosphingolipids and related glycoconjugates. This activity supports lysosomal catabolism and membrane lipid homeostasis through the glycosphingolipid degradation pathway, linking lysosome function to cellular quality control processes such as autophagy and endolysosomal trafficking. Disruption of α-gal A function is classically associated with glycolipid accumulation and lysosomal storage phenotypes, making Gla a relevant target for studying metabolic stress responses and organelle dysfunction. In mouse models, Gla perturbation can be used to interrogate how altered lipid turnover influences inflammation, signaling, and cell-type-specific susceptibility to lysosomal burden.
α-gal A CRISPR/Cas9 KO Plasmid (m2) is a pool of plasmids designed for targeted disruption of the Gla gene in mouse cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the Gla 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 Gla 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 α-gal A protein expression.
This CRISPR knockout system enables efficient generation of Gla-deficient cell models for investigation of α-gal A signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting Gla exon(s) critical for α-gal A 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 Gla genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by α-gal A CRISPR/Cas9 KO Plasmid (m) and α-gal A CRISPR/Cas9 KO Plasmid (m2) target distinct sites within the Gla locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by α-gal A HDR Plasmid (m) and α-gal A HDR Plasmid (m2) contain a puromycin resistance cassette and an RFP reporter flanked by Gla homology arms to support homology-directed repair at defined Gla 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.