Fis1 CRISPR Activation Plasmid (h): sc-401007-ACT

Human FIS1 encodes mitochondrial fission 1 protein (Fis1), an outer mitochondrial membrane adaptor that helps recruit and organize the fission machinery to control mitochondrial division, morphology, and distribution. By coordinating mitochondrial dynamics with bioenergetic state, mitophagy, and apoptotic signaling, Fis1 contributes to cellular stress responses and quality control of the mitochondrial network. Altered FIS1 expression or fission–fusion imbalance has been linked to mitochondrial dysfunction phenotypes observed in neurodegeneration, cardiometabolic disorders, and cancer-associated metabolic rewiring. In experimental systems, modulation of FIS1 is used to probe how mitochondrial architecture influences ROS handling, calcium homeostasis, and innate immune signaling from mitochondria.

Fis1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous FIS1 expression without altering the underlying DNA sequence.

Fis1 CRISPR Activation Plasmid (h) 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.