BIGM103 CRISPR Activation Plasmid (h): sc-403471-ACT

SLC39A8 encodes the human metal ion transporter ZIP8, which regulates cellular uptake of divalent cations such as Mn2+ and Zn2+ and thereby supports metalloenzyme activity, mitochondrial function, and redox homeostasis. By modulating intracellular manganese availability, SLC39A8 influences glycosylation capacity and broader metabolic signaling networks that depend on Mn2+-requiring enzymes. Altered SLC39A8 expression or function has been linked to dysregulated metal homeostasis and downstream effects on inflammatory signaling and cellular stress responses. These connections make SLC39A8 (BIGM103) relevant for mechanistic studies of metal transport, glycan biology, and pathway perturbations in human cell models.

BIGM103 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SLC39A8 expression without altering the underlying DNA sequence.

BIGM103 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SLC39A8 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 SLC39A8 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous BIGM103 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SLC39A8 locus and enabling the study of BIGM103-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of BIGM103 pathway restoration in tumor cells with silenced or reduced SLC39A8 expression.

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