CLIP-170 CRISPR Activation Plasmid (h): sc-401279-ACT

Human CLIP1 encodes CLIP-170, a cytoplasmic plus-end tracking protein that binds microtubule ends and links them to cellular cargos and cortical sites. CLIP-170 coordinates microtubule dynamics, kinetochore capture, and interactions with dynein–dynactin and other +TIP complexes, supporting intracellular transport, spindle organization, and cell polarity. Through these roles, CLIP1 contributes to processes such as mitotic progression and directed migration, with altered microtubule regulation and trafficking networks frequently implicated in tumor cell behavior and neurodevelopmental dysfunction. CLIP-170 is therefore widely studied in pathways governing cytoskeletal remodeling, chromosome segregation, and organelle positioning.

CLIP-170 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CLIP1 expression without altering the underlying DNA sequence.

CLIP-170 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CLIP1 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 CLIP1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous CLIP-170 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CLIP1 locus and enabling the study of CLIP-170-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of CLIP-170 pathway restoration in tumor cells with silenced or reduced CLIP1 expression.

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