BCAP CRISPR Activation Plasmid (h): sc-404405-ACT
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- BCAP CRISPR Activation Plasmid (h) is a synergistic activation mediator (SAM) transcription activation system designed to specifically upregulate gene expression
- BCAP 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 BCAP CRISPR Activation Plasmid (h) and BCAP CRISPR Activation Plasmid (h2) 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 (h) | sc-404405-ACT | 20 µg | $397.00 |
PIK3AP1 encodes BCAP (B cell adaptor for PI3K), a signaling scaffold that couples antigen receptor and innate immune receptor inputs to class I PI3K activation in hematopoietic cells. Through interactions with phosphotyrosine-containing receptors and regulatory components of PI3K, BCAP promotes downstream AKT/mTOR signaling, influences NF-κB and MAPK pathway outputs, and helps shape cytokine production, survival, and differentiation programs. In human immunology, altered PIK3AP1/BCAP signaling has been linked to dysregulated B cell and myeloid responses, contributing to inflammatory phenotypes and immune system dysfunction. Its pathway positioning makes it a useful node for studying receptor-proximal signal integration and feedback control in immune cell activation states.
BCAP CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PIK3AP1 expression without altering the underlying DNA sequence.
BCAP CRISPR Activation Plasmid (h) 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.