α3e Tubulin CRISPR Activation Plasmid (h): sc-400023-ACT

TUBA3E encodes α3e tubulin, a β-tubulin–binding subunit that polymerizes into microtubules to support cytoskeletal architecture, intracellular transport, and the formation and function of the mitotic spindle. As a core component of the microtubule network, α3e tubulin contributes to processes including cell cycle progression, organelle positioning, and neurite outgrowth through dynamic microtubule assembly and remodeling. Tubulin isotypes can modulate microtubule stability and interactions with microtubule-associated proteins and motor complexes, influencing pathways that govern proliferation and cellular polarity. Dysregulated microtubule dynamics and altered tubulin expression patterns are frequently associated with chromosomal instability and aberrant cell division observed in multiple disease contexts, supporting mechanistic studies of cytoskeletal regulation.

α3e Tubulin CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous TUBA3E expression without altering the underlying DNA sequence.

α3e Tubulin CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the TUBA3E 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 TUBA3E transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous α3e Tubulin expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native TUBA3E locus and enabling the study of α3e Tubulin-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of α3e Tubulin pathway restoration in tumor cells with silenced or reduced TUBA3E expression.

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