hDcp1a CRISPR Activation Plasmid (h): sc-402543-ACT

DCP1A encodes hDcp1a, a core component of cytoplasmic processing bodies (P-bodies) that scaffold mRNA decapping and 5′→3′ decay. hDcp1a cooperates with the DCP2 decapping enzyme and decapping activators to coordinate mRNA turnover, translational repression, and RNA quality control, linking stress responses to dynamic changes in gene expression. Through regulation of transcript stability, DCP1A influences pathways such as nonsense-mediated mRNA decay, microRNA-mediated silencing, and stress granule/P-body crosstalk. Dysregulated mRNA decay and P-body biology have been associated with altered proteostasis and aberrant signaling programs observed in cancer and neurodegenerative disease research contexts.

Dcp1a CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous DCP1A expression without altering the underlying DNA sequence.

Dcp1a CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the DCP1A 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 DCP1A transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Dcp1a expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native DCP1A locus and enabling the study of Dcp1a-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Dcp1a pathway restoration in tumor cells with silenced or reduced DCP1A expression.

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