Neu1 CRISPR Activation Plasmid (h): sc-401488-ACT

NEU1 encodes human neuraminidase 1 (Neu1), a lysosomal sialidase that removes terminal sialic acids from glycoproteins and glycolipids, shaping lysosomal catabolism and cell-surface glycan composition. Neu1 functions within the lysosomal multienzyme complex and contributes to turnover and trafficking of sialylated substrates, influencing receptor signaling and endocytic pathways through regulation of protein desialylation. Altered NEU1 activity is linked to disrupted lysosomal homeostasis and aberrant glycosylation patterns that affect inflammatory signaling and cellular adhesion processes. NEU1 dysfunction is associated with lysosomal storage disease phenotypes, making Neu1 a relevant target for studying sialylation-dependent mechanisms in human cells.

Neu1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous NEU1 expression without altering the underlying DNA sequence.

Neu1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the NEU1 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 NEU1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Neu1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native NEU1 locus and enabling the study of Neu1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Neu1 pathway restoration in tumor cells with silenced or reduced NEU1 expression.

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