Fis1 CRISPR Activation Plasmid (m): sc-426020-ACT

Fis1 (fission 1) encodes an outer mitochondrial membrane protein that acts as an adaptor for DRP1-dependent mitochondrial fission, shaping mitochondrial network architecture in response to metabolic state and cellular stress. By coordinating mitochondrial division with organelle distribution, mitophagy, and apoptosis signaling, Fis1 influences bioenergetic homeostasis and reactive oxygen species handling. Dysregulated fission–fusion balance involving Fis1 has been linked to impaired mitochondrial quality control and cellular dysfunction observed across neurodegeneration, cardiometabolic stress, and inflammatory contexts in biomedical research. Mouse Fis1 is therefore widely used to interrogate mitochondrial dynamics pathways and their downstream effects on cell fate decisions.

Fis1 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Fis1 expression without altering the underlying DNA sequence.

Fis1 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Fis1 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 Fis1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Fis1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Fis1 locus and enabling the study of Fis1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Fis1 pathway restoration in tumor cells with silenced or reduced Fis1 expression.

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