Slit2 CRISPR Activation Plasmid (m): sc-423021-ACT

Mouse Slit2 encodes a secreted guidance cue best known for binding Robo receptors to regulate axon pathfinding, neuronal migration, and midline repulsion during development. Beyond the nervous system, SLIT2–ROBO signaling modulates cytoskeletal dynamics, cell adhesion, and directional motility through Rho family GTPases and downstream actin remodeling pathways. Slit2 activity has been implicated in vascular patterning and immune cell trafficking, linking it to processes such as angiogenesis and tissue remodeling. Dysregulated SLIT2/ROBO signaling has been associated with altered invasion and metastatic behavior in cancer models and with aberrant neurodevelopmental phenotypes, supporting its relevance in mechanistic studies of guidance and migration.

Slit2 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Slit2 expression without altering the underlying DNA sequence.

Slit2 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Slit2 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 Slit2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Slit2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Slit2 locus and enabling the study of Slit2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Slit2 pathway restoration in tumor cells with silenced or reduced Slit2 expression.

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