TLE1 CRISPR Activation Plasmid (h): sc-400343-ACT

TLE1 (transducin-like enhancer of split 1) is a Groucho/TLE family transcriptional corepressor that modulates gene expression by interacting with DNA-binding transcription factors and recruiting chromatin-modifying complexes. It is best known for shaping developmental and lineage-specification programs through repression of Wnt/β-catenin target genes and crosstalk with Notch, TGF-β, and other differentiation-associated pathways. By influencing chromatin state and transcriptional output, TLE1 contributes to control of proliferation, cell fate decisions, and tissue homeostasis. Dysregulated TLE1 expression or signaling context is implicated in oncogenic transcriptional programs and is widely studied as a molecular marker and regulatory node in cancer biology.

TLE1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous TLE1 expression without altering the underlying DNA sequence.

TLE1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the TLE1 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 TLE1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous TLE1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native TLE1 locus and enabling the study of TLE1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of TLE1 pathway restoration in tumor cells with silenced or reduced TLE1 expression.

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