N-Shc CRISPR Activation Plasmid (h): sc-401571-ACT

SHC3 encodes the neuronal adaptor protein N-Shc, a member of the Shc family that couples activated receptor tyrosine kinases to intracellular signaling cascades. N-Shc participates in neurotrophin and growth factor signaling by linking receptors such as Trk and EGFR to downstream Ras/MAPK and PI3K/AKT pathways, shaping neuronal differentiation, survival, and synaptic plasticity. Through its PTB and SH2 domains, N-Shc integrates phosphorylation-dependent interactions that influence cytoskeletal dynamics and neurite outgrowth. Altered SHC3 signaling has been associated with dysregulated neural signaling programs studied in neurodevelopmental and neurodegenerative disease models, as well as oncogenic pathway rewiring in select contexts.

N-Shc CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SHC3 expression without altering the underlying DNA sequence.

N-Shc CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SHC3 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 SHC3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous N-Shc expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SHC3 locus and enabling the study of N-Shc-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of N-Shc pathway restoration in tumor cells with silenced or reduced SHC3 expression.

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