D930020B18Rik CRISPR Activation Plasmid (m): sc-432083-ACT

D930020B18Rik encodes a mouse protein that remains incompletely characterized, and available annotations suggest a role in fundamental cellular homeostasis rather than a dedicated, well-mapped signaling module. As an expressed locus, it is frequently considered in transcriptomic and functional genomics studies where modulation of poorly annotated genes can reveal contributions to transcriptional regulation, RNA processing, or protein turnover. Altered expression of uncharacterized RIKEN cDNAs is commonly observed as a secondary feature in disease-relevant contexts such as proliferation, differentiation, and cellular stress responses, supporting its use as a discovery target. Investigating D930020B18Rik can help refine gene network models and identify pathway linkages through phenotypic profiling and multi-omics integration.

D930020B18Rik CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous D930020B18Rik expression without altering the underlying DNA sequence.

D930020B18Rik CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the D930020B18Rik 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 D930020B18Rik transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous D930020B18Rik expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native D930020B18Rik locus and enabling the study of D930020B18Rik-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of D930020B18Rik pathway restoration in tumor cells with silenced or reduced D930020B18Rik expression.

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