GD3 Synthase CRISPR Activation Plasmid (h): sc-404015-ACT
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- GD3 Synthase CRISPR Activation Plasmid (h) is a synergistic activation mediator (SAM) transcription activation system designed to specifically upregulate gene expression
- GD3 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 GD3 Synthase CRISPR Activation Plasmid (h) and GD3 Synthase CRISPR Activation Plasmid (h2) target distinct regulatory regions upstream of the ST8SIA1 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: GD3 Synthase Antibody (B-11): sc-390123
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
GD3 Synthase CRISPR Activation Plasmid (h) | sc-404015-ACT | 20 µg | $397.00 | |||
GD3 Synthase CRISPR Activation Plasmid (h2) | sc-404015-ACT-2 | 20 µg | $397.00 |
ST8SIA1 encodes GD3 synthase, a Golgi-resident sialyltransferase that catalyzes conversion of GM3 to the disialoganglioside GD3, shaping glycosphingolipid composition at the plasma membrane. By modulating ganglioside-dependent membrane microdomains, GD3 synthase influences receptor signaling, cell–cell interactions, and vesicular trafficking processes that integrate with pathways controlling proliferation, migration, and stress responses. Altered ST8SIA1 activity has been linked to changes in glycosylation patterns observed in cancer biology and neurobiology, where ganglioside balance can affect differentiation programs and immune recognition. These properties make ST8SIA1 a useful node for dissecting sialylation-driven regulation of signaling networks and membrane organization.
GD3 Synthase CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ST8SIA1 expression without altering the underlying DNA sequence.
GD3 Synthase CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ST8SIA1 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 ST8SIA1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous GD3 Synthase expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ST8SIA1 locus and enabling the study of GD3 Synthase-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of GD3 Synthase pathway restoration in tumor cells with silenced or reduced ST8SIA1 expression.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.