BIGM103 CRISPR Activation Plasmid (m): sc-426620-ACT

Slc39a8 encodes a ZIP family metal ion transporter implicated in cellular uptake of divalent cations, particularly manganese, supporting metal-dependent enzyme activity and redox homeostasis in mouse tissues. By modulating intracellular manganese availability, BIGM103 influences processes such as glycosylation, mitochondrial function, and inflammatory signaling pathways that depend on metalloenzymes and proper cofactor balance. Perturbation of Slc39a8 expression has been linked to altered immune responses and metabolic phenotypes, making it relevant for studying how micronutrient transport interfaces with cell signaling and systemic physiology. This gene is frequently investigated in models of dysregulated metal homeostasis and complex trait biology where transporter-dependent cofactor supply can reshape downstream pathway activity.

BIGM103 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Slc39a8 expression without altering the underlying DNA sequence.

BIGM103 CRISPR Activation Plasmid (m) 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.