TMEM98 CRISPR Activation Plasmid (h): sc-409870-ACT

TMEM98 encodes a multi-pass transmembrane protein enriched in ocular and neural tissues, where it is implicated in regulating membrane-associated signaling and cellular homeostasis. Human TMEM98 has been linked to processes influencing extracellular matrix organization and developmental patterning, consistent with roles in tissue morphogenesis and cell–cell communication. Genetic studies associate TMEM98 variation with inherited eye phenotypes, including optic nerve and anterior segment abnormalities, supporting its relevance to mechanisms that shape retinal and optic nerve structure. These features make TMEM98 a useful target for studying how membrane proteins tune transcriptional programs and microenvironmental cues in development and disease-associated remodeling.

TMEM98 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous TMEM98 expression without altering the underlying DNA sequence.

TMEM98 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the TMEM98 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 TMEM98 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous TMEM98 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native TMEM98 locus and enabling the study of TMEM98-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of TMEM98 pathway restoration in tumor cells with silenced or reduced TMEM98 expression.

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