G530011O06Rik CRISPR Activation Plasmid (m): sc-437289-ACT

Mouse G530011O06Rik encodes the G530011O06Rik protein, a poorly characterized factor with limited functional annotation in current pathway databases. Available evidence suggests it may participate in fundamental cellular processes such as transcriptional regulation, RNA metabolism, or other context-dependent regulatory programs, making it relevant for systems-level studies of gene network architecture. Because genes in the RIKEN cDNA collection often represent lineage- or stimulus-responsive transcripts, modulating G530011O06Rik expression can help clarify cell state transitions, stress responses, and developmental programs. Defining its endogenous role may inform mechanistic research in disease-relevant models where dysregulated gene expression contributes to pathological phenotypes, without implying clinical utility.

G530011O06Rik CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous G530011O06Rik expression without altering the underlying DNA sequence.

G530011O06Rik CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the G530011O06Rik 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 G530011O06Rik transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous G530011O06Rik expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native G530011O06Rik locus and enabling the study of G530011O06Rik-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of G530011O06Rik pathway restoration in tumor cells with silenced or reduced G530011O06Rik expression.

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