SYNDIG1 CRISPR Activation Plasmid (h): sc-408960-ACT

SYNDIG1 (synapse differentiation induced gene 1) encodes a neuronal transmembrane protein enriched at excitatory synapses, where it contributes to synaptogenesis and the organization of postsynaptic signaling complexes. It is implicated in the regulation of AMPA receptor trafficking and activity-dependent synaptic maturation, linking it to pathways controlling excitatory neurotransmission and neuronal circuit formation. Through its role in synaptic development and plasticity, altered SYNDIG1 expression or function is relevant to studies of neurodevelopmental and neuropsychiatric disease mechanisms characterized by excitatory/inhibitory imbalance and disrupted glutamatergic signaling.

SYNDIG1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SYNDIG1 expression without altering the underlying DNA sequence.

SYNDIG1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SYNDIG1 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 SYNDIG1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous SYNDIG1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SYNDIG1 locus and enabling the study of SYNDIG1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of SYNDIG1 pathway restoration in tumor cells with silenced or reduced SYNDIG1 expression.

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