β8 Tubulin CRISPR Activation Plasmid (h): sc-401438-ACT

TUBB8 encodes β8 tubulin, a primate-specific β-tubulin isotype that heterodimerizes with α-tubulin to form microtubules, supporting cytoskeletal organization, intracellular transport, and chromosome segregation during cell division. Microtubule dynamics regulated by tubulin isotype composition influence spindle assembly, cell polarity, and trafficking pathways that coordinate mitosis and early developmental processes. In humans, TUBB8 variation is linked to defects in oocyte meiotic spindle integrity and early embryonic competence, making it relevant to studies of reproductive biology and cytoskeletal regulation. Altered tubulin network behavior is also broadly informative for modeling stress responses, mitotic errors, and microtubule-associated phenotypes in cell systems.

β8 Tubulin CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous TUBB8 expression without altering the underlying DNA sequence.

β8 Tubulin CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the TUBB8 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 TUBB8 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous β8 Tubulin expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native TUBB8 locus and enabling the study of β8 Tubulin-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of β8 Tubulin pathway restoration in tumor cells with silenced or reduced TUBB8 expression.

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