SPATA18 CRISPR Activation Plasmid (h): sc-407015-ACT

Human SPATA18 encodes a mitochondria-associated factor implicated in mitochondrial quality control and the stress-responsive removal of damaged mitochondria through mitophagy-like processes. It is linked to cellular responses to oxidative stress and maintenance of mitochondrial membrane integrity, connecting SPATA18 activity to broader pathways governing energy metabolism, reactive oxygen species homeostasis, and organelle turnover. Altered regulation of SPATA18 has been reported in contexts of cellular stress and genome maintenance programs, making it relevant to studies of mitochondrial dysfunction and proliferation-associated phenotypes. These functions position SPATA18 as a useful node for investigating how mitochondrial surveillance interfaces with cell fate decisions and disease-associated metabolic remodeling.

SPATA18 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SPATA18 expression without altering the underlying DNA sequence.

SPATA18 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SPATA18 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 SPATA18 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous SPATA18 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SPATA18 locus and enabling the study of SPATA18-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of SPATA18 pathway restoration in tumor cells with silenced or reduced SPATA18 expression.

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