Shank 1 CRISPR Activation Plasmid (h): sc-403064-ACT

SHANK1 encodes Shank 1, a multidomain scaffolding protein enriched at the postsynaptic density of excitatory synapses where it organizes glutamatergic signaling complexes. By linking receptors and channels to actin-regulatory and signaling proteins, Shank 1 supports synapse formation, dendritic spine morphology, and activity-dependent synaptic plasticity. SHANK1 participates in pathways governing postsynaptic architecture and neurotransmission, including assemblies centered on NMDA/AMPA receptor complexes and downstream small GTPase and cytoskeletal remodeling processes. Genetic and functional perturbations of SHANK-family scaffolds are associated with neurodevelopmental and neuropsychiatric phenotypes, making SHANK1 a target for mechanistic studies of synaptic dysfunction.

Shank 1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SHANK1 expression without altering the underlying DNA sequence.

Shank 1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SHANK1 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 SHANK1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Shank 1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SHANK1 locus and enabling the study of Shank 1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Shank 1 pathway restoration in tumor cells with silenced or reduced SHANK1 expression.

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