SAP 145 CRISPR Activation Plasmid (h): sc-403214-ACT

SF3B2 encodes SAP 145, a core component of the SF3b subcomplex within the U2 small nuclear ribonucleoprotein that promotes spliceosome assembly and accurate recognition of the branch point during pre-mRNA splicing. By coordinating splice-site selection and exon definition, SAP 145 supports transcriptome integrity and influences downstream processes including cell-cycle progression, DNA damage responses, and stress-adaptive gene expression programs. Dysregulated spliceosomal factor function can shift isoform balance and alter expression of pathways controlling proliferation and apoptosis, making SF3B2 a useful node for studying mechanisms of aberrant RNA processing in disease-relevant contexts.

SAP 145 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SF3B2 expression without altering the underlying DNA sequence.

SAP 145 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SF3B2 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 SF3B2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous SAP 145 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SF3B2 locus and enabling the study of SAP 145-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of SAP 145 pathway restoration in tumor cells with silenced or reduced SF3B2 expression.

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