FUCA2 CRISPR Activation Plasmid (h): sc-405221-ACT

FUCA2 encodes alpha-L-fucosidase 2, a lysosomal glycosidase involved in the stepwise degradation of fucosylated glycoconjugates, including N-glycans and glycolipids. By regulating terminal fucose removal, FUCA2 contributes to glycan turnover and influences broader lysosome-dependent catabolic processes that interface with cellular quality control and metabolic homeostasis. Altered fucosidase activity and disruptions in fucosylation balance have been associated with changes in cell–cell interactions and immune-related signaling, making FUCA2 relevant for studies of glycometabolism and lysosomal biology. Dysregulated glycosylation pathways are frequently observed in complex disorders, supporting the use of FUCA2 as a mechanistic node in biomarker-oriented and pathway-mapping research.

FUCA2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous FUCA2 expression without altering the underlying DNA sequence.

FUCA2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the FUCA2 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 FUCA2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous FUCA2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native FUCA2 locus and enabling the study of FUCA2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of FUCA2 pathway restoration in tumor cells with silenced or reduced FUCA2 expression.

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