TBL1Y CRISPR Activation Plasmid (h): sc-404389-ACT

TBL1Y encodes a Y-linked transducin beta-like protein that functions as a component of the NCoR/SMRT transcriptional corepressor machinery, helping couple chromatin-associated repression and activation programs with signal-dependent gene regulation. Through interactions with histone-modifying enzymes and transcription factors, TBL1Y-related complexes influence RNA polymerase II transcription, nuclear receptor signaling, and developmental gene expression dynamics. Although less characterized than the paralog TBL1XR1, TBL1 family proteins are broadly implicated in pathways governing cell fate decisions, inflammatory signaling, and oncogenic transcriptional rewiring. Studying TBL1Y supports investigations into sex-linked regulatory networks, transcriptional coregulator biology, and dysregulation of chromatin-dependent gene expression in disease-relevant contexts.

TBL1Y CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous TBL1Y expression without altering the underlying DNA sequence.

TBL1Y CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the TBL1Y 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 TBL1Y transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous TBL1Y expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native TBL1Y locus and enabling the study of TBL1Y-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of TBL1Y pathway restoration in tumor cells with silenced or reduced TBL1Y expression.

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