
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
BUBR1 CRISPR Activation Plasmid (h) | sc-403807-ACT | 20 µg | $397.00 |
BUB1B encodes BUBR1, a core component of the spindle assembly checkpoint that safeguards chromosome segregation by monitoring kinetochore–microtubule attachment and regulating APC/C activity through the mitotic checkpoint complex. BUBR1 contributes to mitotic timing, sister chromatid cohesion, and error correction, thereby maintaining genome stability during cell division. Dysregulated BUB1B/BUBR1 function is associated with chromosomal instability and aneuploidy, features frequently studied in cancer biology and developmental disorders affecting cell cycle control. As a checkpoint regulator, BUBR1 is also used as a mechanistic node for interrogating mitotic stress responses and proliferation-dependent phenotypes.
BUBR1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous BUB1B expression without altering the underlying DNA sequence.
BUBR1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the BUB1B 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 BUB1B transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous BUBR1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native BUB1B locus and enabling the study of BUBR1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of BUBR1 pathway restoration in tumor cells with silenced or reduced BUB1B expression.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.