Freud-1 CRISPR Activation Plasmid (h): sc-408241-ACT

Human CC2D1A encodes Freud-1, a coiled-coil and C2 domain–containing regulatory protein that functions as a DNA-binding transcriptional modulator and signaling scaffold. Freud-1 has been linked to control of serotonergic and dopaminergic gene programs, including regulation of receptor gene promoters, and participates in cellular processes such as transcriptional repression/activation balance, neuronal differentiation, and synaptic signaling. CC2D1A also intersects with membrane-associated signaling pathways, including Akt-related signaling, consistent with roles in cell survival and stress responses. Altered CC2D1A/Freud-1 activity has been associated with neurodevelopmental and neuropsychiatric phenotypes, supporting its relevance in mechanistic studies of neuronal gene regulation.

Freud-1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CC2D1A expression without altering the underlying DNA sequence.

Freud-1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CC2D1A 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 CC2D1A transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Freud-1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CC2D1A locus and enabling the study of Freud-1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Freud-1 pathway restoration in tumor cells with silenced or reduced CC2D1A expression.

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