BUB1 CRISPR Activation Plasmid (h): sc-402006-ACT

BUB1 encodes a serine/threonine kinase that functions as a core component of the spindle assembly checkpoint, coordinating accurate chromosome alignment and segregation during mitosis. It localizes to kinetochores and helps recruit and regulate checkpoint proteins including BUB3, MAD1/2, and CDC20 to control anaphase onset and maintain genomic stability. Through its roles in kinetochore–microtubule attachment and checkpoint signaling, BUB1 links mitotic timing to error correction pathways that prevent aneuploidy. Dysregulated BUB1 expression or activity has been associated with chromosomal instability phenotypes observed across diverse cancer models, making it a useful node for studying mitotic control, proliferation, and genome maintenance mechanisms.

BUB1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous BUB1 expression without altering the underlying DNA sequence.

BUB1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the BUB1 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 BUB1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous BUB1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native BUB1 locus and enabling the study of BUB1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of BUB1 pathway restoration in tumor cells with silenced or reduced BUB1 expression.

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