beta Tubulin CRISPR Activation Plasmid (h): sc-400044-ACT

Human TUBB encodes beta tubulin, a core structural subunit of microtubules that polymerizes with alpha tubulin to build the mitotic spindle and maintain cytoskeletal architecture. Beta tubulin dynamics support chromosome segregation, intracellular transport via kinesin and dynein motors, cell polarity, and cilia-related processes through continuous microtubule remodeling. TUBB activity intersects with cell-cycle regulation and microtubule-dependent signaling that shapes proliferation and migration. Altered tubulin expression or microtubule behavior is broadly relevant to studies of chromosomal instability, neurodevelopmental and neurodegenerative mechanisms, and cytoskeleton-driven changes observed in cancer cell biology.

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

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

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