EB1 CRISPR Activation Plasmid (h): sc-401367-ACT

Human MAPRE1 encodes end-binding protein 1 (EB1), a core microtubule plus-end tracking factor that coordinates microtubule dynamics with cortical capture, mitotic spindle assembly, and chromosome segregation. EB1 interacts with APC and other +TIP proteins to regulate microtubule growth, cell polarity, and intracellular trafficking, linking cytoskeletal remodeling to cell-cycle control. MAPRE1/EB1 dysregulation has been associated with altered mitotic fidelity, aneuploidy, and invasive cell behavior, making it relevant for studying pathways underlying proliferative and migratory phenotypes. Its central role in microtubule-dependent processes also supports mechanistic research in centrosome function and spindle checkpoint regulation.

EB1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous MAPRE1 expression without altering the underlying DNA sequence.

EB1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the MAPRE1 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 MAPRE1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous EB1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native MAPRE1 locus and enabling the study of EB1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of EB1 pathway restoration in tumor cells with silenced or reduced MAPRE1 expression.

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