EHD CRISPR Activation Plasmid (h): sc-407060-ACT

Human EHD1 (Eps15 homology domain-containing protein 1) encodes an ATPase of the EHD protein family that coordinates endocytic membrane remodeling and recycling of internalized receptors back to the plasma membrane. EHD1 regulates trafficking from tubular endosomes and interfaces with Rab-mediated transport, actin dynamics, and phosphoinositide-dependent membrane curvature, influencing receptor availability and signaling duration. Through control of cargo recycling, EHD1 impacts processes such as cell migration, nutrient uptake, and surface expression of adhesion molecules and growth factor receptors. Altered endocytic recycling and receptor trafficking programs involving EHD1 have been linked to phenotypes relevant to proliferative signaling and invasive behavior in cancer biology and to broader dysregulation of membrane transport in complex disease contexts.

EHD1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous EHD1 expression without altering the underlying DNA sequence.

EHD1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the EHD1 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 EHD1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous EHD1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native EHD1 locus and enabling the study of EHD1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of EHD1 pathway restoration in tumor cells with silenced or reduced EHD1 expression.

For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.