Mfn1/Mitofusin 1 CRISPR Activation Plasmid (h): sc-400637-ACT

Human MFN1 encodes Mitofusin 1 (Mfn1), a dynamin-like GTPase embedded in the outer mitochondrial membrane that mediates mitochondrial fusion and shapes mitochondrial network architecture. Mfn1 coordinates with MFN2 and OPA1 to regulate cristae organization, mitochondrial DNA maintenance, and bioenergetic efficiency, linking fusion–fission balance to mitophagy and apoptosis signaling. By controlling organelle connectivity and distribution, MFN1 influences cellular metabolism and stress responses in pathways tied to oxidative phosphorylation and reactive oxygen species homeostasis. Dysregulation of mitochondrial dynamics involving MFN1 has been associated with neurodegeneration, cardiomyocyte dysfunction, and metabolic phenotypes, supporting its relevance in studies of mitochondrial quality control and disease mechanisms.

Mfn1/Mitofusin 1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous MFN1 expression without altering the underlying DNA sequence.

Mfn1/Mitofusin 1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the MFN1 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 MFN1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Mfn1/Mitofusin 1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native MFN1 locus and enabling the study of Mfn1/Mitofusin 1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Mfn1/Mitofusin 1 pathway restoration in tumor cells with silenced or reduced MFN1 expression.

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