Axl CRISPR Activation Plasmid (h): sc-400393-ACT

Human AXL encodes Axl, a receptor tyrosine kinase in the TAM family that is activated by ligands such as GAS6 to regulate cell survival, proliferation, migration, and clearance of apoptotic cells. Axl signaling engages PI3K–AKT, MAPK/ERK, and NF-κB pathway nodes and contributes to epithelial–mesenchymal transition, cytoskeletal remodeling, and modulation of innate immune responses. Dysregulated AXL expression or activity has been associated with invasive tumor phenotypes, altered stromal–immune interactions, and resistance-like signaling states in multiple cancer models. Beyond oncology, Axl-mediated control of inflammatory tone and phagocytic programs links AXL to fibrosis, vascular biology, and infectious disease–relevant immune regulation in experimental systems.

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

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

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