TCF7L2/TCF4 CRISPR Activation Plasmid (h): sc-400607-ACT

TCF7L2 (also known as TCF4) encodes a high-mobility group (HMG) DNA-binding transcription factor that serves as a principal nuclear effector of canonical Wnt/β-catenin signaling. By partnering with β-catenin at TCF/LEF response elements, TCF7L2 regulates gene programs controlling cell fate decisions, proliferation, differentiation, and metabolic homeostasis. In human tissues, TCF7L2 activity influences endocrine and epithelial transcriptional networks and is frequently studied in the context of glucose regulation and developmental patterning. Genetic and expression alterations in TCF7L2 have been associated with metabolic traits and multiple cancers, supporting its broad relevance to disease-linked regulatory circuitry.

TCF7L2/TCF4 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous TCF7L2 expression without altering the underlying DNA sequence.

TCF7L2/TCF4 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the TCF7L2 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 TCF7L2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous TCF7L2/TCF4 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native TCF7L2 locus and enabling the study of TCF7L2/TCF4-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of TCF7L2/TCF4 pathway restoration in tumor cells with silenced or reduced TCF7L2 expression.

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