PRP4 kinase CRISPR Activation Plasmid (m): sc-422432-ACT

Mouse Prpf4b encodes PRP4 kinase, a serine/threonine kinase that couples pre-mRNA splicing control to cell-cycle progression by phosphorylating spliceosomal components and modulating catalytic spliceosome assembly. PRP4 kinase activity influences RNA processing programs that shape transcriptional outputs, with downstream effects on genome stability and stress responses. Dysregulated splicing kinase signaling has been linked to altered proliferation and aberrant transcript isoforms in cancer-related pathways, making Prpf4b a useful node for studying splicing-dependent regulation. In mouse models, perturbing Prpf4b supports mechanistic work on RNA metabolism, checkpoint signaling, and context-specific vulnerabilities in rapidly dividing or differentiation-committed cells.

PRP4 kinase CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Prpf4b expression without altering the underlying DNA sequence.

PRP4 kinase CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Prpf4b 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 Prpf4b transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous PRP4 kinase expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Prpf4b locus and enabling the study of PRP4 kinase-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of PRP4 kinase pathway restoration in tumor cells with silenced or reduced Prpf4b expression.

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