SLC35A2 CRISPR Activation Plasmid (h): sc-408155-ACT

SLC35A2 encodes a Golgi-localized UDP-galactose transporter that imports activated galactose from the cytosol into the Golgi lumen to support galactosylation of N- and O-linked glycans and glycolipids. By controlling nucleotide-sugar availability, SLC35A2 influences glycoprotein maturation, cell-surface receptor function, vesicular trafficking, and secretory pathway homeostasis. Altered SLC35A2 activity has been linked to defects in protein glycosylation with downstream effects on cell signaling and tissue development, and it is frequently studied in the context of congenital disorders of glycosylation and mosaic mutation biology. This gene is therefore relevant for research on glycan-dependent mechanisms that shape adhesion, immune recognition, and membrane protein stability.

SLC35A2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SLC35A2 expression without altering the underlying DNA sequence.

SLC35A2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SLC35A2 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 SLC35A2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous SLC35A2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SLC35A2 locus and enabling the study of SLC35A2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of SLC35A2 pathway restoration in tumor cells with silenced or reduced SLC35A2 expression.

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