Not CRISPR data: the image illustrates antibody quality for visualizing CRISPR experiment results.
Not CRISPR data: the image illustrates antibody quality for visualizing CRISPR experiment results.
twist CRISPR Activation Plasmid (h): sc-400108-ACT
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- twist CRISPR Activation Plasmid (h) is a synergistic activation mediator (SAM) transcription activation system designed to specifically upregulate gene expression
- twist 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 twist CRISPR Activation Plasmid (h) and twist CRISPR Activation Plasmid (h2) target distinct regulatory regions upstream of the TWIST1 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: twist Antibody (Twist2C1a): sc-81417
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
twist CRISPR Activation Plasmid (h) | sc-400108-ACT | 20 µg | $397.00 | |||
twist CRISPR Activation Plasmid (h2) | sc-400108-ACT-2 | 20 µg | $397.00 |
Human TWIST1 encodes twist, a basic helix–loop–helix transcription factor that orchestrates mesenchymal lineage programs and regulates epithelial–mesenchymal transition (EMT) through transcriptional control of cell adhesion, motility, and extracellular matrix remodeling genes. TWIST1 integrates cues from developmental and stress-responsive pathways, including TGF-β, WNT/β-catenin, and PI3K/AKT signaling, to modulate proliferation, differentiation, and survival. Dysregulated TWIST1 activity is linked to altered morphogenesis and aberrant EMT-like states that are frequently studied in tumor progression, invasion, and metastasis models, as well as in fibrosis and stem-like cellular phenotypes. As a nodal regulator of transcriptional plasticity, TWIST1 is widely used to probe gene regulatory networks governing cell state transitions.
twist CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous TWIST1 expression without altering the underlying DNA sequence.
twist CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the TWIST1 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 TWIST1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous twist expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native TWIST1 locus and enabling the study of twist-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of twist pathway restoration in tumor cells with silenced or reduced TWIST1 expression.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.