DAP12 CRISPR Activation Plasmid (h): sc-400916-ACT

TYROBP encodes DNAX-activating protein 12 (DAP12), an ITAM-containing transmembrane adaptor that couples activating receptors on myeloid and lymphoid cells to downstream signaling. DAP12 partners with receptors such as TREM2, SIRPβ1, and certain NK cell receptors to promote phosphorylation of SYK/ZAP70 and engage PI3K, MAPK, and NF-κB pathways that regulate phagocytosis, cytokine production, oxidative burst, and cell survival. In microglia and macrophages, TYROBP-dependent signaling shapes innate immune activation, lipid handling, and clearance of cellular debris, linking it to neuroinflammatory processes and immune dysregulation. Altered TYROBP/DAP12 signaling has been associated with inflammatory and neurodegeneration-related phenotypes, making it a useful target for studying receptor-adaptor signaling networks in human immune cells.

DAP12 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous TYROBP expression without altering the underlying DNA sequence.

DAP12 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the TYROBP 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 TYROBP transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous DAP12 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native TYROBP locus and enabling the study of DAP12-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of DAP12 pathway restoration in tumor cells with silenced or reduced TYROBP expression.

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