Y14 CRISPR Activation Plasmid (h): sc-403255-ACT

RBM8A encodes the human RNA-binding protein Y14, a core component of the exon junction complex that is deposited on mRNA after splicing and coordinates downstream RNA metabolism. Y14 participates in nonsense-mediated mRNA decay, mRNA export, and regulation of translation, thereby influencing transcript quality control and gene expression homeostasis. Through these functions, RBM8A impacts pathways linked to cell-cycle progression, developmental gene regulation, and stress-responsive RNA processing. Dysregulation of exon junction complex activity and RBM8A-associated RNA surveillance has been connected to aberrant splicing and altered transcript stability observed in multiple disease-relevant contexts, including neurodevelopmental and cancer-associated gene expression programs.

Y14 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous RBM8A expression without altering the underlying DNA sequence.

Y14 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the RBM8A 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 RBM8A transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Y14 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native RBM8A locus and enabling the study of Y14-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Y14 pathway restoration in tumor cells with silenced or reduced RBM8A expression.

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