HDMCP CRISPR Activation Plasmid (h): sc-415327-ACT

Human SLC25A47 encodes the mitochondrial inner membrane transporter HDMCP, a member of the solute carrier family implicated in controlling mitochondrial metabolite exchange and respiratory efficiency. HDMCP activity is linked to regulation of oxidative phosphorylation, mitochondrial membrane potential, and broader energy homeostasis programs that coordinate nutrient utilization in metabolically active tissues. Altered expression of mitochondrial carriers, including HDMCP, has been associated with dysregulated lipid and glucose metabolism, oxidative stress, and signaling changes that influence cellular adaptation to energetic demand. As a result, SLC25A47 is of interest for studying mitochondrial bioenergetics, metabolic pathway remodeling, and mechanisms connecting mitochondrial function to disease-relevant phenotypes.

HDMCP CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SLC25A47 expression without altering the underlying DNA sequence.

HDMCP CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SLC25A47 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 SLC25A47 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous HDMCP expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SLC25A47 locus and enabling the study of HDMCP-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of HDMCP pathway restoration in tumor cells with silenced or reduced SLC25A47 expression.

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