DIEXF CRISPR Activation Plasmid (h): sc-411950-ACT

Human DIEXF encodes a conserved RNA-processing factor implicated in ribosome biogenesis and RNA metabolism, with reported roles in pre-rRNA maturation and nucleolar function that support protein synthesis and cell proliferation. DIEXF activity intersects with fundamental gene expression programs by influencing the production and quality control of ribosomal subunits, linking it to cellular growth, stress adaptation, and genome maintenance. Altered regulation of ribosome biogenesis components is frequently observed in proliferative and stress-related states, making DIEXF a useful node for studying pathways that couple nucleolar homeostasis to cell-cycle control and viability. In biomedical research, DIEXF is therefore relevant for interrogating how perturbations in RNA processing and translational capacity reshape cellular phenotypes.

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

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

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