SNRPA CRISPR Activation Plasmid (h): sc-404128-ACT

SNRPA encodes the U1 small nuclear ribonucleoprotein A, a core component of the U1 snRNP that recognizes 5′ splice sites and initiates spliceosome assembly during pre-mRNA splicing. Through its role in splice-site selection and coupling of transcription to RNA processing, SNRPA helps maintain proteome integrity and regulated gene expression programs. Perturbation of spliceosomal proteins such as SNRPA can reshape alternative splicing landscapes, influencing pathways including RNA surveillance and cell-cycle control. Dysregulated splicing machinery is frequently observed across tumor biology and neurodegenerative contexts, making SNRPA a useful node for studying RNA-processing stress and spliceosome dependency in human cells.

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

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

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