TMEM43 CRISPR Activation Plasmid (h): sc-402764-ACT

TMEM43 (transmembrane protein 43), also known as LUMA, is an inner nuclear membrane protein that associates with the nuclear lamina and contributes to nuclear envelope organization, mechanotransduction, and maintenance of nuclear architecture. By interacting with lamins and other envelope components, TMEM43 influences chromatin positioning and gene regulatory programs linked to cell differentiation and stress responses. Altered TMEM43 function has been connected to cardiac and neuromuscular phenotypes, including arrhythmogenic cardiomyopathy, highlighting its relevance for studying nuclear envelope–associated disease mechanisms. In human cell models, TMEM43 perturbation is often used to probe links between nuclear structure, transcriptional control, and cell-state stability.

TMEM43 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous TMEM43 expression without altering the underlying DNA sequence.

TMEM43 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the TMEM43 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 TMEM43 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous TMEM43 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native TMEM43 locus and enabling the study of TMEM43-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of TMEM43 pathway restoration in tumor cells with silenced or reduced TMEM43 expression.

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