CD77 synthase CRISPR Activation Plasmid (h): sc-404828-ACT
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- CD77 synthase CRISPR Activation Plasmid (h) is a synergistic activation mediator (SAM) transcription activation system designed to specifically upregulate gene expression
- CD77 synthase 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 CD77 synthase CRISPR Activation Plasmid (h) and CD77 synthase CRISPR Activation Plasmid (h2) target distinct regulatory regions upstream of the A4GALT transcriptional start site. One or both designs may be available
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
CD77 synthase CRISPR Activation Plasmid (h) | sc-404828-ACT | 20 µg | $397.00 |
A4GALT encodes CD77 synthase, a Golgi-localized glycosyltransferase that catalyzes addition of galactose to lactosylceramide to generate globotriaosylceramide (Gb3/CD77), a key neutral glycosphingolipid. By shaping glycosphingolipid biosynthesis, CD77 synthase influences membrane microdomain organization, protein trafficking, and cell–cell recognition processes relevant to immune and epithelial cell biology. Altered A4GALT activity and CD77/Gb3 abundance have been associated with changes in differentiation state and susceptibility to glycan-dependent interactions, including pathways implicated in B-cell biology and tumor-associated glycosylation phenotypes. As a result, A4GALT is frequently studied in the context of glycosphingolipid metabolism, glycome remodeling, and cell-surface antigen regulation.
CD77 synthase CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous A4GALT expression without altering the underlying DNA sequence.
CD77 synthase CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the A4GALT 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 A4GALT transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous CD77 synthase expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native A4GALT locus and enabling the study of CD77 synthase-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of CD77 synthase pathway restoration in tumor cells with silenced or reduced A4GALT expression.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.