PRP19 CRISPR Activation Plasmid (h): sc-404130-ACT

PRPF19 encodes the human PRP19 protein, a core component of the PRP19/CDC5L complex that couples pre-mRNA splicing with genome maintenance. PRP19 functions as a scaffold and E3 ubiquitin ligase in spliceosome activation and has been implicated in coordinating transcription, RNA processing, and DNA damage response signaling to preserve replication fidelity. Through these roles, PRPF19 activity influences cell-cycle progression and proteostasis pathways that shape cellular stress tolerance. Dysregulation of PRPF19-associated splicing and DNA repair programs is frequently studied in the context of oncogenic transformation, neurodegeneration, and other disorders where RNA metabolism and genome stability are perturbed.

PRP19 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PRPF19 expression without altering the underlying DNA sequence.

PRP19 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PRPF19 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 PRPF19 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous PRP19 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PRPF19 locus and enabling the study of PRP19-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of PRP19 pathway restoration in tumor cells with silenced or reduced PRPF19 expression.

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