U1 SnRNP 70 CRISPR Activation Plasmid (h): sc-401829-ACT

SNRNP70 encodes U1 SnRNP 70, a core component of the U1 small nuclear ribonucleoprotein that recognizes 5′ splice sites and initiates spliceosome assembly during pre-mRNA splicing. Through its interactions with U1 snRNA and additional spliceosomal factors, U1 SnRNP 70 supports accurate exon definition, co-transcriptional RNA processing, and global regulation of gene expression programs. Disruption of spliceosome function and altered splice-site selection are recurrent features of cellular stress responses and oncogenic transcriptional rewiring, making SNRNP70 a useful node for studying RNA processing control. Aberrant splicing patterns linked to spliceosomal perturbation are also relevant to neurodegeneration and other diseases where RNA metabolism is compromised, motivating mechanistic studies of U1-associated pathways.

U1 SnRNP 70 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SNRNP70 expression without altering the underlying DNA sequence.

U1 SnRNP 70 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SNRNP70 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 SNRNP70 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous U1 SnRNP 70 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SNRNP70 locus and enabling the study of U1 SnRNP 70-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of U1 SnRNP 70 pathway restoration in tumor cells with silenced or reduced SNRNP70 expression.

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