CREST CRISPR Activation Plasmid (h): sc-403134-ACT

SS18L1 (CREST) encodes a neuron-enriched chromatin-associated factor that interfaces with transcriptional regulation and activity-dependent gene expression programs. CREST cooperates with chromatin remodeling machinery, including SWI/SNF-related complexes, to modulate promoter and enhancer states that influence neuronal differentiation, neurite outgrowth, and synaptic function. Through these epigenetic mechanisms, SS18L1 links neuronal signaling to long-term changes in transcriptional networks and cellular identity. Dysregulation of CREST-associated pathways has been investigated in the context of neurodevelopmental and neurodegenerative disease-relevant processes, including altered synaptic homeostasis and stress-responsive transcription.

CREST CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SS18L1 expression without altering the underlying DNA sequence.

CREST CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SS18L1 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 SS18L1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous CREST expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SS18L1 locus and enabling the study of CREST-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of CREST pathway restoration in tumor cells with silenced or reduced SS18L1 expression.

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