Rap 1A CRISPR Activation Plasmid (h): sc-400572-ACT
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Rap 1A CRISPR Activation Plasmid (h) is a synergistic activation mediator (SAM) transcription activation system designed to specifically upregulate gene expression
- Rap 1A 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 Rap 1A CRISPR Activation Plasmid (h) and Rap 1A CRISPR Activation Plasmid (h2) target distinct regulatory regions upstream of the RAP1A 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: Rap 1A Antibody (C-10): sc-373968
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Rap 1A CRISPR Activation Plasmid (h) | sc-400572-ACT | 20 µg | $397.00 | |||
Rap 1A CRISPR Activation Plasmid (h2) | sc-400572-ACT-2 | 20 µg | $397.00 |
RAP1A encodes Rap 1A, a Ras-family small GTPase that cycles between GDP- and GTP-bound states to coordinate signal transduction at the plasma membrane and endomembranes. Active Rap 1A regulates integrin inside-out signaling, cell–cell and cell–matrix adhesion, and actin cytoskeleton remodeling, influencing migration, polarity, and barrier function. Through effectors such as RAF kinases, RIAM, and RalGDS, Rap 1A intersects with MAPK signaling and pathways controlling vesicle trafficking and junctional dynamics. Dysregulated RAP1A activity has been linked to oncogenic signaling contexts and altered invasive behavior, supporting its use as a mechanistic node in studies of tumor progression, vascular biology, and immune cell trafficking.
Rap 1A CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous RAP1A expression without altering the underlying DNA sequence.
Rap 1A CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the RAP1A 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 RAP1A transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Rap 1A expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native RAP1A locus and enabling the study of Rap 1A-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Rap 1A pathway restoration in tumor cells with silenced or reduced RAP1A expression.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.