
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
AGAP1 CRISPR Activation Plasmid (h) | sc-409504-ACT | 20 µg | $397.00 |
AGAP1 (ArfGAP with GTPase domain, ankyrin repeat and PH domain 1) is an ARF GTPase-activating protein that coordinates membrane trafficking and cytoskeletal dynamics by regulating ARF-dependent vesicle formation and endosomal transport. Through its PH and ankyrin repeat domains, AGAP1 links phosphoinositide signaling to adaptor-mediated sorting, influencing receptor recycling, neurite outgrowth, and actin remodeling. Altered AGAP1 expression or function has been associated with dysregulated endocytic pathways and synaptic processes, making it relevant to studies of neurodevelopmental phenotypes and cellular homeostasis. Its roles at endosomes and the plasma membrane also intersect with signaling networks that depend on receptor availability and spatially restricted phosphoinositide pools.
AGAP1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous AGAP1 expression without altering the underlying DNA sequence.
AGAP1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the AGAP1 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 AGAP1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous AGAP1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native AGAP1 locus and enabling the study of AGAP1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of AGAP1 pathway restoration in tumor cells with silenced or reduced AGAP1 expression.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.