CLASP1 CRISPR Activation Plasmid (h): sc-404219-ACT

CLASP1 (cytoplasmic linker associated protein 1) is a microtubule plus-end tracking protein that stabilizes and organizes microtubule networks at the cell cortex and kinetochores, coordinating spindle assembly, chromosome alignment, and faithful segregation during mitosis. Through interactions with EB proteins, CLIP proteins, and kinetochore-associated factors, CLASP1 supports microtubule rescue and dynamic instability that are essential for cell polarity, intracellular trafficking, and directed migration. Perturbation of CLASP1-regulated microtubule dynamics can promote mitotic errors and aneuploidy, linking its function to pathways broadly relevant to proliferative control and genome stability. Accordingly, CLASP1 is frequently investigated in studies of cytoskeletal regulation, cell-cycle progression, and mechanisms that connect microtubule behavior to cellular transformation and stress responses.

CLASP1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CLASP1 expression without altering the underlying DNA sequence.

CLASP1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CLASP1 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 CLASP1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous CLASP1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CLASP1 locus and enabling the study of CLASP1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of CLASP1 pathway restoration in tumor cells with silenced or reduced CLASP1 expression.

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