CLIP4 CRISPR Activation Plasmid (h): sc-413231-ACT

CLIP4 (CAP-Gly domain containing linker protein 4) encodes a cytoskeletal-associated factor implicated in regulating microtubule dynamics and intracellular organization through interactions with tubulin and microtubule plus-end networks. By influencing microtubule stability and spatial coordination of the cytoskeleton, CLIP4 can affect processes such as vesicle trafficking, cell polarity, and mitotic progression. Altered expression or dysregulated cytoskeletal control is commonly linked to aberrant cell migration and proliferative phenotypes, making CLIP4 a relevant target for mechanistic studies in cancer biology and other disorders involving cytoskeletal remodeling. CLIP4 research also supports investigation of pathway cross-talk between microtubule regulation and signaling programs that tune cell shape and adhesion.

CLIP4 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CLIP4 expression without altering the underlying DNA sequence.

CLIP4 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CLIP4 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 CLIP4 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous CLIP4 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CLIP4 locus and enabling the study of CLIP4-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of CLIP4 pathway restoration in tumor cells with silenced or reduced CLIP4 expression.

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