TFIID/TATA Binding Protein/TBP CRISPR Activation Plasmid (h): sc-400579-ACT

TBP encodes the TATA-binding protein, a conserved core component of the TFIID complex that recognizes promoter elements and nucleates assembly of the RNA polymerase II preinitiation complex. By binding the TATA box and coordinating recruitment of general transcription factors, TBP supports basal transcription and influences promoter selection, chromatin engagement, and transcriptional initiation dynamics. TBP also contributes to transcriptional programs driven by Pol I and Pol III, linking it to ribosome biogenesis, tRNA synthesis, and global proteostasis. Altered TBP dosage or functional perturbation has been associated with dysregulated gene expression and neurodegenerative phenotypes, making it a useful node for studying transcriptional homeostasis in disease-relevant models.

TFIID/TATA Binding Protein/TBP CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous TBP expression without altering the underlying DNA sequence.

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

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