A33 CRISPR Activation Plasmid (h): sc-404457-ACT
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- A33 CRISPR Activation Plasmid (h) is a synergistic activation mediator (SAM) transcription activation system designed to specifically upregulate gene expression
- A33 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 A33 CRISPR Activation Plasmid (h) and A33 CRISPR Activation Plasmid (h2) target distinct regulatory regions upstream of the GPA33 transcriptional start site. One or both designs may be available
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
A33 CRISPR Activation Plasmid (h) | sc-404457-ACT | 20 µg | $397.00 |
GPA33 encodes the human A33 glycoprotein, a single-pass transmembrane cell-surface molecule implicated in epithelial differentiation and maintenance of intercellular adhesion. A33 is enriched in intestinal epithelium and is used as a lineage-associated marker in studies of gastrointestinal cell identity, membrane trafficking, and epithelial barrier biology. Its expression patterns and altered regulation are frequently evaluated in colorectal and other gastrointestinal disease research to interrogate changes in tumor-associated epithelial programs. GPA33/A33 therefore provides a tractable target for dissecting surface antigen biology and epithelial signaling states in relevant cellular models.
A33 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous GPA33 expression without altering the underlying DNA sequence.
A33 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the GPA33 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 GPA33 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous A33 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native GPA33 locus and enabling the study of A33-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of A33 pathway restoration in tumor cells with silenced or reduced GPA33 expression.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.