MREG CRISPR Activation Plasmid (h): sc-405725-ACT

Human MREG (melanoregulin) encodes a small, membrane-associated protein implicated in melanosome biogenesis and pigment granule transport, influencing the maturation and distribution of lysosome-related organelles. MREG participates in intracellular trafficking processes that intersect with endosomal–lysosomal dynamics and cytoskeletal transport, shaping organelle positioning and cargo handling. Through its roles in pigmentation biology and organelle homeostasis, altered MREG activity is relevant to studies of pigmentary phenotypes and broader defects in vesicle transport pathways. These functions make MREG a useful node for dissecting mechanisms of organelle maturation, cargo sorting, and cellular responses to trafficking perturbations.

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

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

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