Tyro3 CRISPR Activation Plasmid (m): sc-423567-ACT

Tyro3 encodes a receptor tyrosine kinase of the TAM family that regulates cell survival, proliferation, and cytoskeletal remodeling in response to ligands such as GAS6 and Protein S. In mouse tissues, TYRO3 signaling interfaces with PI3K–AKT, MAPK/ERK, and JAK/STAT pathways, shaping processes that include phagocytic clearance of apoptotic cells and modulation of innate immune tone. Through crosstalk with AXL and MERTK, Tyro3 contributes to homeostatic control of inflammatory signaling and can influence cell state transitions in diverse microenvironments. Dysregulated TAM receptor activity has been associated with aberrant immune regulation and oncogenic signaling contexts, making Tyro3 a useful node for mechanistic studies in neurobiology, immunology, and cancer biology models.

Tyro3 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Tyro3 expression without altering the underlying DNA sequence.

Tyro3 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Tyro3 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 Tyro3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Tyro3 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Tyro3 locus and enabling the study of Tyro3-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Tyro3 pathway restoration in tumor cells with silenced or reduced Tyro3 expression.

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