BCAP CRISPR Activation Plasmid (m): sc-429913-ACT
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- BCAP CRISPR Activation Plasmid (m) is a synergistic activation mediator (SAM) transcription activation system designed to specifically upregulate gene expression
- BCAP 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 BCAP CRISPR Activation Plasmid (m) and BCAP CRISPR Activation Plasmid (m2) target distinct regulatory regions upstream of the Pik3ap1 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: BCAP Antibody (E-5): sc-515498
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
BCAP CRISPR Activation Plasmid (m) | sc-429913-ACT | 20 µg | $397.00 | |||
BCAP CRISPR Activation Plasmid (m2) | sc-429913-ACT-2 | 20 µg | $397.00 |
Mouse Pik3ap1 encodes BCAP (B cell adaptor for PI3K), a cytoplasmic adaptor that links immunoreceptor signaling to phosphoinositide 3-kinase activation. BCAP couples upstream cues from the B cell receptor and innate immune receptors to PI3K–AKT signaling, shaping downstream transcriptional programs that influence lymphocyte activation, survival, and metabolic reprogramming. Through these roles, Pik3ap1 is used to study signal integration across immune pathways, including crosstalk with NF-κB and MAPK-dependent responses. Dysregulation of PI3K pathway wiring and adaptor-mediated signal propagation is relevant to models of immune dysfunction and inflammation, supporting mechanistic research in immunology and systems signaling.
BCAP CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Pik3ap1 expression without altering the underlying DNA sequence.
BCAP CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Pik3ap1 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 Pik3ap1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous BCAP expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Pik3ap1 locus and enabling the study of BCAP-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of BCAP pathway restoration in tumor cells with silenced or reduced Pik3ap1 expression.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.