DMXL2 CRISPR Activation Plasmid (h): sc-417505-ACT

DMXL2 (Dmx-like 2) encodes a large WD-repeat–containing scaffold protein implicated in vesicle trafficking and regulated exocytosis, supporting membrane dynamics at secretory and synaptic compartments. By coordinating protein–protein interactions within endomembrane pathways, DMXL2 is linked to neuronal and endocrine cell function, including processes that couple vesicle maturation, docking, and release. Altered DMXL2 expression or function has been associated with neurodevelopmental phenotypes and reproductive/endocrine dysregulation, making it relevant for studying secretory pathway control in human cells. As a broadly expressed regulator of membrane trafficking, DMXL2 is frequently examined in models of synaptic biology, hormone secretion, and cellular homeostasis.

DMXL2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous DMXL2 expression without altering the underlying DNA sequence.

DMXL2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the DMXL2 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 DMXL2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous DMXL2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native DMXL2 locus and enabling the study of DMXL2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of DMXL2 pathway restoration in tumor cells with silenced or reduced DMXL2 expression.

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