POD-1 CRISPR Activation Plasmid (h): sc-402890-ACT

TCF21 encodes the basic helix–loop–helix transcription factor POD-1, a lineage-determining regulator that coordinates mesenchymal differentiation programs and organ development, including epicardial, renal, and gonadal stromal lineages. POD-1 modulates transcriptional networks that intersect with epithelial–mesenchymal transitions, extracellular matrix remodeling, and cell-cycle control through context-dependent interactions with other bHLH factors and developmental signaling pathways. In adult tissues, altered TCF21 expression is linked to changes in fibroblast activation states and stromal remodeling, impacting gene programs relevant to tissue homeostasis. Dysregulation of TCF21/POD-1 has been associated with cancer-relevant transcriptional reprogramming and fibrotic or remodeling phenotypes, making it useful for studying stromal biology and differentiation-associated gene networks.

POD-1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous TCF21 expression without altering the underlying DNA sequence.

POD-1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the TCF21 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 TCF21 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous POD-1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native TCF21 locus and enabling the study of POD-1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of POD-1 pathway restoration in tumor cells with silenced or reduced TCF21 expression.

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