FucT-IX CRISPR Activation Plasmid (h): sc-416354-ACT

FUT9 encodes α1,3-fucosyltransferase IX (FucT-IX), a Golgi-resident glycosyltransferase that catalyzes terminal fucosylation steps in the biosynthesis of Lewis-type antigens, including Lewis X (CD15). By shaping cell-surface and secreted glycan structures, FucT-IX influences glycan-dependent recognition events involved in cell adhesion, migration, and immune cell interactions. FUT9 activity intersects with broader glycosylation networks that modulate receptor signaling, developmental programs, and lineage-specific differentiation, particularly in neural and hematopoietic contexts. Altered fucosylation patterns and FUT9 dysregulation have been associated with changes in tumor cell behavior and inflammatory microenvironments, making FUT9 a useful target for mechanistic studies of glyco-epitopes in disease biology.

FucT-IX CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous FUT9 expression without altering the underlying DNA sequence.

FucT-IX CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the FUT9 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 FUT9 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous FucT-IX expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native FUT9 locus and enabling the study of FucT-IX-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of FucT-IX pathway restoration in tumor cells with silenced or reduced FUT9 expression.

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