TPPP3 CRISPR Activation Plasmid (h): sc-405113-ACT

TPPP3 (tubulin polymerization promoting protein family member 3) is a microtubule-associated protein implicated in cytoskeletal organization and dynamics, influencing processes such as cell shape, intracellular transport, and cell-cycle progression. By modulating tubulin polymerization and microtubule stability, TPPP3 can impact pathways linked to epithelial differentiation, motility, and stress responses. Altered TPPP3 expression has been reported in multiple disease contexts, including carcinomas where changes in cytoskeletal regulation correlate with invasion and proliferation phenotypes. These properties make TPPP3 a useful target for mechanistic studies of microtubule-dependent signaling and phenotypic plasticity in human cells.

TPPP3 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous TPPP3 expression without altering the underlying DNA sequence.

TPPP3 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the TPPP3 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 TPPP3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous TPPP3 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native TPPP3 locus and enabling the study of TPPP3-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of TPPP3 pathway restoration in tumor cells with silenced or reduced TPPP3 expression.

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