Tra-2 CRISPR Activation Plasmid (h): sc-404739-ACT

Human TRA2B encodes Tra-2, an RNA-binding splicing regulator that recognizes exonic splicing enhancer elements and modulates exon inclusion across diverse pre-mRNA targets. Tra-2 participates in spliceosome-associated networks that shape alternative splicing programs controlling cell-cycle progression, differentiation, and stress-responsive transcriptional outputs. Dysregulated TRA2B expression or activity has been linked to widespread splicing alterations observed in cancer and neurodevelopmental contexts, where isoform imbalance can remodel signaling and RNA metabolism. As a central node in post-transcriptional gene regulation, TRA2B is frequently studied to connect sequence-specific splicing control with pathway rewiring and disease-associated transcriptome changes.

Tra-2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous TRA2B expression without altering the underlying DNA sequence.

Tra-2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the TRA2B 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 TRA2B transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Tra-2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native TRA2B locus and enabling the study of Tra-2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Tra-2 pathway restoration in tumor cells with silenced or reduced TRA2B expression.

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