SF2/ASF CRISPR Activation Plasmid (h): sc-400737-ACT

Human SRSF1 encodes the serine/arginine-rich splicing factor SF2/ASF, a core component of the spliceosome that regulates constitutive and alternative pre-mRNA splicing, mRNA export, and translation. SF2/ASF helps shape isoform selection across pathways controlling cell-cycle progression, apoptosis, and stress responses, and it can influence signaling networks such as PI3K–AKT and MAPK through splicing-dependent rewiring of pathway components. Altered SRSF1 expression or activity is linked to widespread transcriptome changes, genomic instability, and dysregulated proteome output observed in multiple disease-relevant contexts, including cancer biology and neurodegeneration. As a prototypical splicing regulator, SRSF1 is frequently studied for its role in exon definition, RNA-binding specificity, and coupling between transcription and splicing.

SF2/ASF CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SRSF1 expression without altering the underlying DNA sequence.

SF2/ASF CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SRSF1 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 SRSF1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous SF2/ASF expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SRSF1 locus and enabling the study of SF2/ASF-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of SF2/ASF pathway restoration in tumor cells with silenced or reduced SRSF1 expression.

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