Not CRISPR data: the image illustrates antibody quality for visualizing CRISPR experiment results.
Not CRISPR data: the image illustrates antibody quality for visualizing CRISPR experiment results.
BSEP CRISPR Activation Plasmid (h): sc-400742-ACT
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- BSEP CRISPR Activation Plasmid (h) is a synergistic activation mediator (SAM) transcription activation system designed to specifically upregulate gene expression
- BSEP 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 BSEP CRISPR Activation Plasmid (h) and BSEP CRISPR Activation Plasmid (h2) target distinct regulatory regions upstream of the ABCB11 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: BSEP Antibody (F-6): sc-74500
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
BSEP CRISPR Activation Plasmid (h) | sc-400742-ACT | 20 µg | $397.00 | |||
BSEP CRISPR Activation Plasmid (h2) | sc-400742-ACT-2 | 20 µg | $397.00 |
ABCB11 encodes the human bile salt export pump (BSEP), an ATP-binding cassette transporter localized to the canalicular membrane of hepatocytes that mediates ATP-dependent efflux of bile acids into bile. This transport activity is central to bile acid homeostasis and enterohepatic circulation, influencing hepatic lipid handling and detoxification processes. ABCB11 function is integrated with bile acid–sensing pathways, including FXR-regulated transcriptional networks that coordinate bile acid synthesis and transport. Dysregulated or reduced BSEP activity is linked to cholestatic phenotypes and hepatocellular stress driven by intracellular bile acid accumulation, making ABCB11 a key target for studying bile formation and bile acid–mediated signaling.
BSEP CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ABCB11 expression without altering the underlying DNA sequence.
BSEP CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ABCB11 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 ABCB11 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous BSEP expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ABCB11 locus and enabling the study of BSEP-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of BSEP pathway restoration in tumor cells with silenced or reduced ABCB11 expression.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.