FUT2 CRISPR Activation Plasmid (h2): sc-409559-ACT-2

Human FUT2 encodes α(1,2)-fucosyltransferase 2 (FUT2), a Golgi-resident glycosyltransferase that catalyzes terminal fucosylation of glycans to generate H type 1 and related fucosylated epitopes on mucosal surfaces and secreted glycoproteins. By controlling secretor status, FUT2 modulates glycan-dependent processes including ABO antigen presentation in secretions, epithelial barrier biology, and host–microbe interactions that shape mucosal immunity and microbiome composition. Genetic variation or loss of FUT2 activity is associated with altered susceptibility to infection and inflammatory phenotypes, and has been linked to traits such as inflammatory bowel disease risk, norovirus susceptibility, and shifts in microbial community structure. FUT2 gene editing and functional perturbation are used to dissect glycosylation pathways, map lectin and pathogen binding determinants, and model how mucosal fucosylation influences immune signaling and epithelial physiology in human cell systems.

FUT2 CRISPR Activation Plasmid (h2) provides a targeted, non-destructive approach to upregulating endogenous FUT2 expression without altering the underlying DNA sequence.

FUT2 CRISPR Activation Plasmid (h2) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the FUT2 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 FUT2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous FUT2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native FUT2 locus and enabling the study of FUT2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of FUT2 pathway restoration in tumor cells with silenced or reduced FUT2 expression.

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