C14orf102 CRISPR Activation Plasmid (h): sc-409988-ACT

NRDE2 (also annotated as C14orf102) encodes a conserved nuclear factor implicated in RNA-mediated gene regulation and surveillance pathways that help maintain genome integrity. Studies in model systems link NRDE2 to heterochromatin-associated transcriptional silencing and coordination of RNA processing with the DNA damage response, supporting roles in cell cycle control and stress adaptation. In human cells, NRDE2 has been associated with nuclear RNA metabolism and chromatin-linked regulation of transcription, processes frequently perturbed in proliferative and genome instability phenotypes. Dysregulation of these pathways is relevant to mechanistic studies of oncogenic transformation and other conditions where defective RNA processing and DNA repair contribute to disease biology.

NRDE-2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous NRDE2 expression without altering the underlying DNA sequence.

NRDE-2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the NRDE2 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 NRDE2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous NRDE-2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native NRDE2 locus and enabling the study of NRDE-2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of NRDE-2 pathway restoration in tumor cells with silenced or reduced NRDE2 expression.

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