Dynamin II CRISPR Activation Plasmid (h): sc-401037-ACT

DNM2 encodes human Dynamin II, a large GTPase that catalyzes membrane fission during clathrin-mediated endocytosis and other vesicular trafficking events. Dynamin II coordinates with actin remodeling, BAR-domain proteins, and adaptor complexes to regulate receptor internalization, synaptic vesicle recycling, and endosomal sorting, thereby shaping signaling outputs and membrane homeostasis. Through its roles in membrane dynamics, DNM2 influences processes such as cytokinesis and organelle distribution, linking endocytic control to broader cell physiology. Altered DNM2 activity or regulation is associated with neuromuscular and neurodevelopmental phenotypes and is frequently investigated for its impact on receptor trafficking and downstream pathway modulation.

Dynamin II CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous DNM2 expression without altering the underlying DNA sequence.

Dynamin II CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the DNM2 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 DNM2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Dynamin II expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native DNM2 locus and enabling the study of Dynamin II-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Dynamin II pathway restoration in tumor cells with silenced or reduced DNM2 expression.

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