TMEM119 CRISPR Activation Plasmid (h): sc-404363-ACT

TMEM119 encodes a type I transmembrane protein enriched in microglia and other myeloid-lineage contexts, where it is widely used as a marker of microglial identity and maturation state. Although its mechanistic roles are still being clarified, TMEM119 expression tracks with programs that shape immune surveillance, cellular homeostasis, and inflammatory signaling in the central nervous system. Altered TMEM119-positive microglial states have been associated with neuroinflammatory and neurodegenerative disease research frameworks, including studies of microglial activation, tissue remodeling, and interactions with neurons and astrocytes. As a membrane-associated protein, TMEM119 is also relevant for investigating cell-surface–linked regulatory networks and lineage-specific transcriptional control.

TMEM119 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous TMEM119 expression without altering the underlying DNA sequence.

TMEM119 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the TMEM119 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 TMEM119 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous TMEM119 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native TMEM119 locus and enabling the study of TMEM119-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of TMEM119 pathway restoration in tumor cells with silenced or reduced TMEM119 expression.

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