GM2/GD2 synthase CRISPR/Cas9 KO Plasmid (m): sc-420474

Overview

B4galnt1 encodes GM2/GD2 synthase, a Golgi-localized glycosyltransferase that transfers N-acetylgalactosamine to ganglioside precursors to generate GM2 and GD2, shaping the composition of glycosphingolipids in cellular membranes. By controlling ganglioside abundance, it influences lipid raft organization, cell–cell interactions, and signaling processes important for neuronal development and synaptic function. In mouse tissues, B4galnt1 activity contributes to myelin-associated glycolipid patterns and neuroimmune homeostasis through regulation of glycolipid-mediated recognition events. Dysregulation of ganglioside biosynthesis is linked to neurological phenotypes and altered cell-surface antigen landscapes, making B4galnt1 a key target for mechanistic studies in neurobiology and membrane biology.

GM2/GD2 synthase CRISPR/Cas9 KO Plasmid (m) is a pool of plasmids designed for targeted disruption of the B4galnt1 gene in mouse cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the B4galnt1 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 B4galnt1 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 GM2/GD2 synthase protein expression.

This CRISPR knockout system enables efficient generation of B4galnt1-deficient cell models for investigation of GM2/GD2 synthase signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.

Key Features

• sgRNAs targeting B4galnt1 exon(s) critical for GM2/GD2 synthase 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 B4galnt1 genomic sites to improve knockout efficiency
• Compatible with delivery by transfection

Design Variants

CRISPRs +/- HDRs

• gRNAs encoded by GM2/GD2 synthase CRISPR/Cas9 KO Plasmid (m) and GM2/GD2 synthase CRISPR/Cas9 KO Plasmid (m2) target distinct sites within the B4galnt1 locus. One or both targeting designs may be available. See Related Products for availability.
• HDR donor constructs encoded by GM2/GD2 synthase HDR Plasmid (m) and GM2/GD2 synthase HDR Plasmid (m2) contain a puromycin resistance cassette and an RFP reporter flanked by B4galnt1 homology arms to support homology-directed repair at defined B4galnt1 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.