DRP1 CRISPR Activation Plasmid (h): sc-400459-ACT

DNM1L encodes dynamin-related protein 1 (DRP1), a large GTPase that orchestrates mitochondrial and peroxisomal fission by assembling on organelle membranes and constricting them in coordination with adaptor proteins such as MFF, MID49/51, and FIS1. Through control of organelle morphology, DRP1 influences bioenergetics, mitophagy, apoptosis signaling, and redistribution of mitochondria during cell division and neuronal transport. DRP1 activity is integrated with stress-response pathways and is regulated by phosphorylation, SUMOylation, ubiquitination, and S-nitrosylation, linking cellular signaling to mitochondrial dynamics. Dysregulated DNM1L/DRP1 function is implicated in neurodevelopmental and neurodegenerative phenotypes, cardiometabolic dysfunction, and cancer-associated metabolic remodeling, making it a key node for mechanistic studies of mitochondrial homeostasis.

DRP1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous DNM1L expression without altering the underlying DNA sequence.

DRP1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the DNM1L 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 DNM1L transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous DRP1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native DNM1L locus and enabling the study of DRP1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of DRP1 pathway restoration in tumor cells with silenced or reduced DNM1L expression.

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