MTMR8/9 CRISPR Activation Plasmid (h): sc-413097-ACT

MTMR9 encodes a myotubularin family protein that participates in phosphoinositide signaling through interactions with catalytically active myotubularins, influencing the cellular balance of phosphatidylinositol 3-phosphate and phosphatidylinositol (3,5)-bisphosphate. Through these lipid-dependent pathways, MTMR8/9-associated complexes are implicated in endosomal trafficking, membrane remodeling, and downstream regulation of nutrient-sensing and autophagy-related processes. MTMR9 has been linked to metabolic phenotypes and insulin-related signaling contexts in genetic and functional studies, and altered expression has been reported in disease-associated datasets including cancer. These features make MTMR9 a useful target for interrogating phosphoinositide-dependent control of vesicle dynamics and cell-state regulation.

MTMR8/9 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous MTMR9 expression without altering the underlying DNA sequence.

MTMR8/9 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the MTMR9 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 MTMR9 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous MTMR8/9 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native MTMR9 locus and enabling the study of MTMR8/9-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of MTMR8/9 pathway restoration in tumor cells with silenced or reduced MTMR9 expression.

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