Shc CRISPR Activation Plasmid (h): sc-400914-ACT

SHC1 encodes the Shc adaptor protein, a central signal transducer that links activated receptor tyrosine kinases and cytokine receptors to downstream cascades including RAS–MAPK/ERK and PI3K–AKT. Through its PTB and SH2 domains, Shc coordinates phosphorylation-dependent assembly of signaling complexes that regulate proliferation, differentiation, survival, and cellular stress responses. SHC1-dependent signaling also interfaces with insulin/IGF pathways and influences mitochondrial and oxidative stress signaling, contributing to context-dependent changes in cell growth and apoptosis. Dysregulated SHC1/Shc signaling has been associated with aberrant growth-factor pathway activity in cancer biology and with altered signaling dynamics in metabolic and neurodegeneration-related research models.

Shc CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SHC1 expression without altering the underlying DNA sequence.

Shc CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SHC1 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 SHC1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Shc expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SHC1 locus and enabling the study of Shc-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Shc pathway restoration in tumor cells with silenced or reduced SHC1 expression.

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