RBM25 CRISPR Activation Plasmid (h): sc-407825-ACT

Human RBM25 encodes an RNA-binding splicing regulator that associates with the spliceosome to influence alternative pre-mRNA processing, exon selection, and mRNA maturation. Through control of transcript isoform balance, RBM25 contributes to RNA quality control and downstream programs affecting cell-cycle progression, DNA damage responses, and stress-adaptive gene expression. Dysregulated RBM25 activity has been linked to aberrant splicing patterns and altered expression of apoptosis- and signaling-related transcripts observed across multiple disease-relevant contexts, including cancer biology. As a splicing factor with broad transcriptome impact, RBM25 is frequently studied to map regulatory networks connecting RNA processing to phenotypic outcomes.

RBM25 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous RBM25 expression without altering the underlying DNA sequence.

RBM25 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the RBM25 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 RBM25 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous RBM25 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native RBM25 locus and enabling the study of RBM25-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of RBM25 pathway restoration in tumor cells with silenced or reduced RBM25 expression.

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