FucT-VIII CRISPR Activation Plasmid (h): sc-402957-ACT
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- FucT-VIII CRISPR Activation Plasmid (h) is a synergistic activation mediator (SAM) transcription activation system designed to specifically upregulate gene expression
- FucT-VIII CRISPR Activation Plasmid (h) consists of three plasmids at a 1:1:1 mass ratio: a plasmid encoding the deactivated Cas9 (dCas9) nuclease (D10A and N863A) fused to the transactivation domain VP64, and a blasticidin resistance gene; a plasmid encoding the MS2-p65-HSF1 fusion protein, and a hygromycin resistance gene; a plasmid encoding a target-specific 20 nt guide RNA fused to two MS2 RNA aptamers, and a puromycin resistance gene
- The resulting SAM complex binds to a site-specific region approximately 200-250 nt upstream of the transcriptional start site and provides robust recruitment of transcription factors for highly efficient gene activation
- gRNAs encoded by FucT-VIII CRISPR Activation Plasmid (h) and FucT-VIII CRISPR Activation Plasmid (h2) target distinct regulatory regions upstream of the FUT8 transcriptional start site. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: FucT-VIII Antibody (B-10): sc-271244
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
FucT-VIII CRISPR Activation Plasmid (h) | sc-402957-ACT | 20 µg | $397.00 | |||
FucT-VIII CRISPR Activation Plasmid (h2) | sc-402957-ACT-2 | 20 µg | $397.00 |
FUT8 encodes alpha-(1,6)-fucosyltransferase (FucT-VIII), the Golgi-resident enzyme that catalyzes core fucosylation of N-linked glycans on glycoproteins. This modification shapes protein folding, secretion, receptor–ligand interactions, and downstream signaling across pathways influenced by glycosylation state, including growth factor and immune receptor networks. Altered FUT8 activity is linked to changes in cell adhesion and migration and has been associated with dysregulated glycoprotein remodeling observed in cancer biology, inflammation, and congenital disorders of glycosylation. As a result, FUT8 is widely studied in glycobiology, secreted protein quality control, and mechanisms that couple Golgi glycan processing to signal transduction.
FucT-VIII CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous FUT8 expression without altering the underlying DNA sequence.
FucT-VIII CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the FUT8 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 FUT8 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous FucT-VIII expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native FUT8 locus and enabling the study of FucT-VIII-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of FucT-VIII pathway restoration in tumor cells with silenced or reduced FUT8 expression.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.