RBM38 CRISPR Activation Plasmid (h): sc-403488-ACT

RBM38 (RNA binding motif protein 38) is a human RNA-binding protein that modulates post-transcriptional gene regulation by controlling mRNA stability, localization, and translation. It is implicated in cell-cycle control and stress-response programs through sequence-specific interactions with target transcripts, influencing pathways linked to proliferation and differentiation. RBM38 activity intersects with RNA processing networks and can shape p53-associated regulatory circuits by tuning expression of factors involved in checkpoint responses. Dysregulation of RBM38 expression has been reported in multiple disease-relevant contexts, making it a useful node for studying RNA-centric mechanisms that affect cellular phenotype.

RBM38 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous RBM38 expression without altering the underlying DNA sequence.

RBM38 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the RBM38 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 RBM38 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous RBM38 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native RBM38 locus and enabling the study of RBM38-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of RBM38 pathway restoration in tumor cells with silenced or reduced RBM38 expression.

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