DND1 CRISPR Activation Plasmid (h): sc-404161-ACT

Human DND1 (dead end protein homolog 1) is an RNA-binding protein that regulates post-transcriptional gene expression by recognizing specific sequence motifs within target mRNAs and modulating their stability and translation. It is best known for roles in germ cell development and maintenance of cellular identity, including coordination of RNA fate decisions that influence proliferation, differentiation, and survival programs. DND1 participates in networks that intersect with microRNA-mediated regulation by limiting access of microRNAs to selected transcripts, thereby shaping proteomic outputs during development. Dysregulation of DND1-dependent RNA control has been associated with altered germline biology and tumor-related phenotypes in model systems, making it relevant for studies of oncogenic reprogramming, cell fate transitions, and RNA regulatory circuits.

DND1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous DND1 expression without altering the underlying DNA sequence.

DND1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the DND1 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 DND1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous DND1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native DND1 locus and enabling the study of DND1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of DND1 pathway restoration in tumor cells with silenced or reduced DND1 expression.

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