TRRAP CRISPR Activation Plasmid (h): sc-402748-ACT

TRRAP (transformation/transcription domain-associated protein) is a large adaptor protein that functions as an essential cofactor for multiple histone acetyltransferase complexes, including SAGA and TIP60, linking sequence-specific transcription factors to chromatin remodeling. By coordinating acetylation-dependent regulation of gene expression, TRRAP influences cell cycle progression, DNA damage responses, and differentiation programs. It participates in pathways governed by MYC and E2F transcriptional networks and supports genome integrity through chromatin-mediated control of repair and replication-associated stress. Dysregulated TRRAP activity and altered complex assembly have been associated with oncogenic transcriptional states and neurodevelopmental phenotypes, making it a relevant target for mechanistic studies of epigenetic regulation.

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

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

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