PI 3-kinase p85α CRISPR Activation Plasmid (m): sc-422233-ACT
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- PI 3-kinase p85α CRISPR Activation Plasmid (m) is a synergistic activation mediator (SAM) transcription activation system designed to specifically upregulate gene expression
- PI 3-kinase p85α 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 PI 3-kinase p85α CRISPR Activation Plasmid (m) and PI 3-kinase p85α CRISPR Activation Plasmid (m2) target distinct regulatory regions upstream of the Pik3r1 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: PI 3-kinase p85α Antibody (B-9): sc-1637
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
PI 3-kinase p85α CRISPR Activation Plasmid (m) | sc-422233-ACT | 20 µg | $397.00 | |||
PI 3-kinase p85α CRISPR Activation Plasmid (m2) | sc-422233-ACT-2 | 20 µg | $397.00 |
Pik3r1 encodes the p85α regulatory subunit of class IA phosphoinositide 3-kinase (PI3K), which stabilizes and recruits p110 catalytic subunits to activated receptor tyrosine kinases and adaptor proteins. Through control of PI(3,4,5)P3 production, PI 3-kinase p85α modulates AKT/mTOR signaling to regulate cell growth, metabolism, survival, and insulin receptor signaling. In mouse models, altered Pik3r1 activity influences immune cell development and function, glucose homeostasis, and responses to growth factor stimulation. Dysregulated PI3K pathway signaling is broadly implicated in oncogenic processes and metabolic disease mechanisms, making Pik3r1 a key node for pathway interrogation.
PI 3-kinase p85α CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Pik3r1 expression without altering the underlying DNA sequence.
PI 3-kinase p85α CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Pik3r1 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 Pik3r1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous PI 3-kinase p85α expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Pik3r1 locus and enabling the study of PI 3-kinase p85α-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of PI 3-kinase p85α pathway restoration in tumor cells with silenced or reduced Pik3r1 expression.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.