hnRNP D0 CRISPR Activation Plasmid (h): sc-401911-ACT

HNRNPD encodes hnRNP D0 (AUF1), an RNA-binding protein that recognizes AU-rich elements in 3′ UTRs and modulates mRNA turnover and translation. By controlling the stability of transcripts involved in inflammation, stress responses, cell-cycle progression, and apoptosis, hnRNP D0 helps shape post-transcriptional gene regulatory networks and proteostasis. It participates in ribonucleoprotein complex assembly and influences alternative RNA processing, linking it to signaling programs such as cytokine/NF-κB-associated transcriptional outputs and cellular senescence. Altered hnRNP D0 expression or RNA-binding activity has been associated with dysregulated cytokine production and oncogenic phenotypes, making it a useful node for studying disease-relevant RNA metabolism.

hnRNP D0 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous HNRNPD expression without altering the underlying DNA sequence.

hnRNP D0 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the HNRNPD 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 HNRNPD transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous hnRNP D0 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native HNRNPD locus and enabling the study of hnRNP D0-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of hnRNP D0 pathway restoration in tumor cells with silenced or reduced HNRNPD expression.

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