TIAR CRISPR Activation Plasmid (h): sc-401247-ACT

Human TIAL1 encodes TIAR, an RNA-binding protein that recognizes U-rich elements to regulate pre-mRNA splicing, mRNA stability, and translation. TIAR is a core component of stress granules and participates in post-transcriptional control programs that reshape gene expression during cellular stress, innate immune signaling, and apoptosis. Through modulation of cytokine and growth-related transcripts, TIAR helps coordinate RNA metabolism with signaling pathways governing proliferation and inflammatory responses. Dysregulated TIAR activity and stress granule dynamics have been linked to neurodegeneration, autoimmunity, and cancer-relevant phenotypes, making TIAL1 a useful node for studying RNA regulatory networks and disease-associated transcriptomic shifts.

TIAR CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous TIAL1 expression without altering the underlying DNA sequence.

TIAR CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the TIAL1 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 TIAL1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous TIAR expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native TIAL1 locus and enabling the study of TIAR-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of TIAR pathway restoration in tumor cells with silenced or reduced TIAL1 expression.

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