Ganglioside sialidase CRISPR Activation Plasmid (h): sc-404848-ACT

NEU3 encodes a human plasma membrane–associated ganglioside sialidase that removes terminal sialic acids from gangliosides and related glycosphingolipids, reshaping cell-surface glycan composition. By modulating ganglioside-dependent membrane microdomains, NEU3 can influence receptor tyrosine kinase signaling, cell adhesion and migration, and downstream MAPK/PI3K-linked pathways that respond to changes in lipid–protein organization. Altered sialidase activity impacts glycosphingolipid metabolism and inflammatory signaling networks, making NEU3 a useful node for studying how glycan remodeling regulates cell communication. Dysregulated NEU3 expression has been reported in contexts of aberrant growth and tissue remodeling, supporting its relevance for mechanistic studies of disease-associated signaling and membrane biology.

Ganglioside sialidase CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous NEU3 expression without altering the underlying DNA sequence.

Ganglioside sialidase CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the NEU3 locus in human cell lines. The system is built around a catalytically inactive Cas9 (dCas9) carrying two inactivating mutations (D10A and N863A) that eliminate nuclease activity while preserving DNA binding. This dCas9 is fused to VP64, a potent transcriptional activator, and is co-expressed with a blasticidin resistance gene for selection. The second plasmid encodes the MS2-p65-HSF1 fusion protein, a secondary activator complex that works in concert with dCas9-VP64, alongside a hygromycin resistance gene. The third plasmid encodes a target-specific 20 nt sgRNA fused to two MS2 RNA aptamers that recruit the MS2-p65-HSF1 complex to the activation site, accompanied by a puromycin resistance gene. The three plasmids are delivered at a 1:1:1 mass ratio for balanced expression of all system components.

Once assembled at the target locus, the SAM complex binds within approximately 200 bp upstream of the NEU3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Ganglioside sialidase expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native NEU3 locus and enabling the study of Ganglioside sialidase-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Ganglioside sialidase pathway restoration in tumor cells with silenced or reduced NEU3 expression.

For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.