MerTK CRISPR Activation Plasmid (h): sc-401629-ACT

Human MERTK encodes the receptor tyrosine kinase MerTK, a TAM family member that recognizes phosphatidylserine on apoptotic cells via ligands such as GAS6 and PROS1 to promote efferocytosis and resolution of inflammation. MerTK signaling engages PI3K–AKT, MAPK/ERK, and STAT pathways to coordinate macrophage and microglial homeostatic programs, dampen innate immune activation, and influence cell survival and migration. In the tumor microenvironment, MerTK-dependent clearance and immunoregulatory signaling can shape cytokine outputs and antigen-presenting cell function, while in the nervous system it contributes to microglial phagocytic activity and tissue remodeling. Dysregulated MERTK expression or signaling has been linked to altered inflammatory tone and aberrant phagocytosis across cancer biology and neuroinflammation-focused research contexts.

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

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

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