TSHZ3 CRISPR Activation Plasmid (h): sc-410072-ACT

TSHZ3 (teashirt zinc finger homeobox 3) encodes a nuclear transcription factor characterized by C2H2 zinc-finger domains that regulate gene expression programs during development and cell-type specification. In human tissues, TSHZ3 contributes to transcriptional control of differentiation and patterning processes, integrating with broader gene regulatory networks that shape lineage identity and organogenesis. Dysregulation of TSHZ3 expression or function has been linked to neurodevelopmental phenotypes and altered circuit formation, making it relevant for studies of transcriptional control in neuronal and mesenchymal lineages. As a DNA-binding regulator, TSHZ3 is commonly investigated in pathways influencing morphogenesis, synaptic development, and cell fate decisions.

TSHZ3 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous TSHZ3 expression without altering the underlying DNA sequence.

TSHZ3 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the TSHZ3 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 TSHZ3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous TSHZ3 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native TSHZ3 locus and enabling the study of TSHZ3-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of TSHZ3 pathway restoration in tumor cells with silenced or reduced TSHZ3 expression.

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