LONP1 CRISPR Activation Plasmid (h): sc-411273-ACT

LONP1 encodes the mitochondrial matrix AAA+ Lon protease, a central component of mitochondrial protein quality control that couples ATP-dependent unfolding to proteolysis of damaged or misfolded proteins. By regulating turnover of respiratory chain subunits and mitochondrial nucleoid-associated factors, LONP1 supports oxidative phosphorylation, mitochondrial DNA maintenance, and the integrated mitochondrial stress response. LONP1 activity intersects with pathways governing proteostasis, ROS homeostasis, and apoptosis, linking mitochondrial dysfunction to altered metabolism and cell fate decisions. Dysregulated LONP1 expression or function has been associated with mitochondrial proteopathy phenotypes and is frequently studied in the context of neurodegeneration, cancer cell metabolic remodeling, and inherited mitochondrial disease mechanisms.

LONP1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous LONP1 expression without altering the underlying DNA sequence.

LONP1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the LONP1 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 LONP1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous LONP1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native LONP1 locus and enabling the study of LONP1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of LONP1 pathway restoration in tumor cells with silenced or reduced LONP1 expression.

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