MTMR2 CRISPR Activation Plasmid (m): sc-429507-ACT

Mtmr2 encodes MTMR2, a phosphoinositide 3-phosphatase that dephosphorylates PI3P and PI(3,5)P2 to regulate endosomal trafficking, membrane remodeling, and phosphoinositide-dependent signaling. In mouse cells, MTMR2 contributes to control of vesicle dynamics and cytoskeletal organization that influence receptor turnover and intracellular transport. Altered MTMR2 activity perturbs endolysosomal homeostasis and myelin-related cellular programs, linking MTMR2-associated pathways to peripheral neuropathy phenotypes. These functions make Mtmr2 a useful node for studying phosphoinositide metabolism and its impact on membrane trafficking networks in neural and non-neural systems.

MTMR2 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Mtmr2 expression without altering the underlying DNA sequence.

MTMR2 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Mtmr2 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 Mtmr2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous MTMR2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Mtmr2 locus and enabling the study of MTMR2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of MTMR2 pathway restoration in tumor cells with silenced or reduced Mtmr2 expression.

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