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.
Apaf-1 CRISPR Activation Plasmid (h): sc-401291-ACT
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Apaf-1 CRISPR Activation Plasmid (h) is a synergistic activation mediator (SAM) transcription activation system designed to specifically upregulate gene expression
- Apaf-1 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 Apaf-1 CRISPR Activation Plasmid (h) and Apaf-1 CRISPR Activation Plasmid (h2) target distinct regulatory regions upstream of the APAF1 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: Apaf-1 Antibody (5E11): sc-65891
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Apaf-1 CRISPR Activation Plasmid (h) | sc-401291-ACT | 20 µg | $397.00 |
APAF1 encodes apoptotic protease activating factor 1 (Apaf-1), a central scaffold in the intrinsic mitochondrial apoptosis pathway. Following cytochrome c release and dATP/ATP binding, Apaf-1 oligomerizes into the apoptosome to recruit and activate initiator caspase-9, propagating downstream caspase cascades that regulate programmed cell death. This node integrates cellular stress signals, mitochondrial outer membrane permeabilization, and checkpoint pathways that govern tissue homeostasis and developmental cell turnover. Dysregulated APAF1 activity has been associated with altered apoptotic competence in cancer biology and neurodegeneration-related models, making it relevant for mechanistic studies of cell fate control.
Apaf-1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous APAF1 expression without altering the underlying DNA sequence.
Apaf-1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the APAF1 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 APAF1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Apaf-1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native APAF1 locus and enabling the study of Apaf-1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Apaf-1 pathway restoration in tumor cells with silenced or reduced APAF1 expression.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.