SCG10 CRISPR Activation Plasmid (m): sc-422814-ACT

Stmn2 encodes SCG10, a neuron-enriched stathmin-family phosphoprotein that regulates microtubule dynamics by binding tubulin and promoting microtubule destabilization during neurite initiation, axon outgrowth, and growth cone remodeling in mouse cells. SCG10 is transported to neurites and growth cones, linking cytoskeletal remodeling to intracellular trafficking and activity-dependent neuronal plasticity. Through its control of microtubule turnover, SCG10 influences processes such as axonal regeneration, synaptic connectivity, and neuronal differentiation programs. Altered STMN2/SCG10 regulation is frequently studied in the context of neurodegeneration and axonopathy, including pathways implicated in motor neuron vulnerability and impaired axonal maintenance.

SCG10 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Stmn2 expression without altering the underlying DNA sequence.

SCG10 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Stmn2 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 Stmn2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous SCG10 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Stmn2 locus and enabling the study of SCG10-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of SCG10 pathway restoration in tumor cells with silenced or reduced Stmn2 expression.

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