α3d Tubulin CRISPR Activation Plasmid (h): sc-400024-ACT

TUBA3D encodes human α3d Tubulin, a core component of microtubules that supports cytoskeletal architecture, intracellular trafficking, and mitotic spindle assembly. As part of the tubulin polymerization cycle, α/β-tubulin heterodimers enable dynamic instability required for cell division, neuronal process outgrowth, and polarized transport. Microtubule remodeling interfaces with pathways controlling centrosome function, chromosome segregation, and cilia-related processes, linking tubulin regulation to genome stability and cell fate decisions. Dysregulated microtubule dynamics and altered tubulin expression patterns are frequently associated with proliferative and neurodevelopmental phenotypes, and are broadly relevant to mechanisms underlying chromosomal instability in cancer models.

α3d Tubulin CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous TUBA3D expression without altering the underlying DNA sequence.

α3d Tubulin CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the TUBA3D 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 TUBA3D transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous α3d Tubulin expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native TUBA3D locus and enabling the study of α3d Tubulin-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of α3d Tubulin pathway restoration in tumor cells with silenced or reduced TUBA3D expression.

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