SLC35B4 CRISPR Activation Plasmid (h): sc-413728-ACT

SLC35B4 encodes a Golgi/ER-associated nucleotide-sugar transporter that mediates uptake of UDP-xylose and UDP-N-acetylglucosamine into the secretory pathway, supplying substrates for glycosylation reactions. By regulating nucleotide-sugar availability, SLC35B4 influences glycan assembly on proteins and lipids, shaping membrane trafficking, receptor signaling, and extracellular matrix interactions. Altered nucleotide-sugar transport and downstream glycosylation states are linked to metabolic adaptation and stress responses, making SLC35B4 a useful node for studying glycometabolism and secretory pathway homeostasis. Dysregulated glycosylation programs have been implicated across cancer biology and inflammatory phenotypes, supporting investigation of SLC35B4 in disease-relevant cell models.

SLC35B4 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SLC35B4 expression without altering the underlying DNA sequence.

SLC35B4 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SLC35B4 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 SLC35B4 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous SLC35B4 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SLC35B4 locus and enabling the study of SLC35B4-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of SLC35B4 pathway restoration in tumor cells with silenced or reduced SLC35B4 expression.

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