RBM10 CRISPR Activation Plasmid (h): sc-418527-ACT

RBM10 encodes an RNA-binding protein that regulates alternative splicing, mRNA stability, and spliceosome assembly, shaping transcript isoform choice across diverse gene programs. RBM10-dependent splicing influences cell-cycle progression, apoptosis, and differentiation, and it interfaces with post-transcriptional control of signaling outputs such as Notch pathway components and other growth-regulatory transcripts. Dysregulated RBM10 activity or mutation has been associated with aberrant RNA processing phenotypes observed in multiple cancer contexts and congenital developmental disorders, making it a useful node for studying how splicing perturbations remodel cellular states. As a nuclear regulator of RNA metabolism, RBM10 is frequently investigated for its impact on transcriptome-wide exon usage, gene expression networks, and stress-responsive RNA processing.

RBM10 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous RBM10 expression without altering the underlying DNA sequence.

RBM10 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the RBM10 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 RBM10 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous RBM10 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native RBM10 locus and enabling the study of RBM10-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of RBM10 pathway restoration in tumor cells with silenced or reduced RBM10 expression.

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