Dapper1 CRISPR Activation Plasmid (h): sc-405361-ACT

Human DACT1 encodes Dapper1, a cytoplasmic scaffold/adaptor protein that modulates Dishevelled-dependent signaling and helps coordinate Wnt pathway output during embryonic development and tissue homeostasis. Dapper1 has been linked to regulation of planar cell polarity and β-catenin–dependent transcription, influencing processes such as cell polarity, migration, and differentiation. Altered DACT1 expression or pathway context has been associated with dysregulated Wnt signaling observed across multiple disease settings, including developmental abnormalities and cancer-related phenotypes. These attributes make DACT1 a useful node for dissecting Wnt network dynamics and cross-talk with other morphogen and cytoskeletal pathways.

Dapper1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous DACT1 expression without altering the underlying DNA sequence.

Dapper1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the DACT1 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 DACT1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Dapper1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native DACT1 locus and enabling the study of Dapper1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Dapper1 pathway restoration in tumor cells with silenced or reduced DACT1 expression.

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