GAP1m CRISPR Activation Plasmid (m): sc-431130-ACT
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- GAP1m CRISPR Activation Plasmid (m) is a synergistic activation mediator (SAM) transcription activation system designed to specifically upregulate gene expression
- GAP1m CRISPR Activation Plasmid (m) 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 GAP1m CRISPR Activation Plasmid (m) and GAP1m CRISPR Activation Plasmid (m2) target distinct regulatory regions upstream of the Rasa2 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: GAP1m Antibody (15): sc-135916
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
GAP1m CRISPR Activation Plasmid (m) | sc-431130-ACT | 20 µg | $397.00 |
Rasa2 encodes the mouse Ras GTPase-activating protein GAP1m, a negative regulator of Ras signaling that accelerates GTP hydrolysis to restrain Ras activity. By limiting downstream RAF–MEK–ERK and PI3K–AKT pathway output, GAP1m influences proliferation, differentiation, and cytoskeletal dynamics in diverse cell types. Altered Rasa2 expression or function can shift signaling thresholds that control growth-factor responsiveness and stress adaptation, making it relevant to studies of oncogenic Ras pathway deregulation and developmental phenotypes. In immune and stromal contexts, modulation of Ras attenuation by Rasa2 is also linked to changes in activation state and tissue homeostasis.
GAP1m CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Rasa2 expression without altering the underlying DNA sequence.
GAP1m CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Rasa2 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 Rasa2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous GAP1m expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Rasa2 locus and enabling the study of GAP1m-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of GAP1m pathway restoration in tumor cells with silenced or reduced Rasa2 expression.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.