TMEM192 CRISPR Activation Plasmid (h): sc-415021-ACT

Human TMEM192 encodes a lysosome-associated transmembrane protein implicated in endolysosomal organization and vesicular trafficking. Its localization to lysosomal membranes supports roles in lysosome integrity, cargo processing, and coordination of autophagy–lysosome pathways that maintain cellular proteostasis. Because lysosomal function influences innate immune signaling, metabolic adaptation, and stress responses, altered TMEM192 activity is relevant to research on neurodegeneration, cancer cell survival, and inflammatory phenotypes where lysosome-dependent processes are frequently perturbed. Modulating TMEM192 expression provides a tractable approach to interrogate lysosome-centric regulation of degradation pathways and signaling crosstalk.

TMEM192 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous TMEM192 expression without altering the underlying DNA sequence.

TMEM192 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the TMEM192 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 TMEM192 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous TMEM192 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native TMEM192 locus and enabling the study of TMEM192-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of TMEM192 pathway restoration in tumor cells with silenced or reduced TMEM192 expression.

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