TREM-1 CRISPR Activation Plasmid (h): sc-417344-ACT

TREM1 encodes triggering receptor expressed on myeloid cells 1 (TREM-1), an immunoreceptor prominently expressed on neutrophils and monocytes/macrophages that amplifies inflammatory signaling in response to microbial products and danger-associated cues. Upon ligand engagement and coupling to the adaptor TYROBP/DAP12, TREM-1 activates SYK-dependent pathways that converge on NF-κB and MAPK signaling, promoting cytokine and chemokine production and shaping innate immune effector functions. This axis intersects with Toll-like receptor signaling and inflammasome-associated processes, influencing leukocyte activation, migration, and tissue inflammatory tone. Dysregulated TREM-1 activity has been linked to exaggerated myeloid inflammation in infectious and inflammatory disease contexts, making it a useful target for dissecting regulation of innate immune responses.

TREM-1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous TREM1 expression without altering the underlying DNA sequence.

TREM-1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the TREM1 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 TREM1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous TREM-1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native TREM1 locus and enabling the study of TREM-1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of TREM-1 pathway restoration in tumor cells with silenced or reduced TREM1 expression.

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