RBM7 CRISPR Activation Plasmid (h): sc-408808-ACT

Human RBM7 encodes an RNA-binding protein that functions as a core component of the nuclear exosome targeting (NEXT) complex, partnering with ZCCHC8 and SKIV2L2/MTR4 to channel specific nuclear RNAs to the RNA exosome for processing and decay. RBM7 preferentially recognizes U-rich elements and contributes to RNA surveillance by promoting turnover of promoter-upstream transcripts, enhancer RNAs, and other short-lived noncoding RNAs, thereby shaping transcriptional output and maintaining nuclear RNA homeostasis. Through coupling of RNA quality control with transcription and chromatin-associated RNA metabolism, RBM7 influences genome stability and stress-responsive gene expression programs. Dysregulation of nuclear RNA decay pathways involving RBM7 has been associated with altered gene regulatory networks and has relevance to studies of cancer-associated transcriptional remodeling and neurodevelopmental phenotypes linked to RNA processing defects.

RBM7 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous RBM7 expression without altering the underlying DNA sequence.

RBM7 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the RBM7 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 RBM7 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous RBM7 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native RBM7 locus and enabling the study of RBM7-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of RBM7 pathway restoration in tumor cells with silenced or reduced RBM7 expression.

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