SRm300 CRISPR Activation Plasmid (h): sc-403447-ACT

SRRM2 encodes SRm300, a large serine/arginine-rich nuclear protein that functions as a scaffold within the spliceosome and regulates pre-mRNA splicing decisions. SRm300 associates with SR proteins and other splicing factors in nuclear speckles to coordinate exon definition, alternative splicing, and coupling of transcription with RNA processing. Through these activities, SRRM2 contributes to transcript isoform diversity and proteome regulation in pathways governing cell cycle progression, DNA damage responses, and cellular differentiation. Dysregulated splicing programs and altered SRRM2-associated spliceosomal dynamics have been linked to disease-relevant phenotypes in cancer and neurodegenerative research contexts, where aberrant isoform usage can influence signaling outputs.

SRm300 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SRRM2 expression without altering the underlying DNA sequence.

SRm300 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SRRM2 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 SRRM2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous SRm300 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SRRM2 locus and enabling the study of SRm300-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of SRm300 pathway restoration in tumor cells with silenced or reduced SRRM2 expression.

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