TIA-1 CRISPR Activation Plasmid (h): sc-400212-ACT

TIA1 encodes TIA-1, an RNA-binding protein that recognizes U-rich sequences and coordinates post-transcriptional gene regulation through alternative splicing, mRNA stabilization, and translational repression. TIA-1 is a key component of stress granules and links cellular stress responses to changes in transcript fate, influencing processes such as apoptosis, innate immune signaling, and inflammatory gene expression. Through its roles in RNA metabolism and stress-adaptive programs, altered TIA-1 activity has been associated with dysregulated cytokine pathways and aberrant RNA processing observed in cancer and neurodegeneration research contexts. These functions make TIA1 a useful node for studying how stress signaling reshapes gene expression networks at the RNA level.

TIA-1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous TIA1 expression without altering the underlying DNA sequence.

TIA-1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the TIA1 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 TIA1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous TIA-1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native TIA1 locus and enabling the study of TIA-1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of TIA-1 pathway restoration in tumor cells with silenced or reduced TIA1 expression.

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