Clathrin Heavy Chain/CLTC CRISPR Activation Plasmid (h): sc-400863-ACT

CLTC encodes clathrin heavy chain, a core structural component of clathrin-coated pits and vesicles that coordinates membrane curvature and cargo selection during clathrin-mediated endocytosis. Through interactions with adaptor protein complexes and accessory factors, CLTC regulates receptor internalization, synaptic vesicle recycling, endosomal trafficking, and trans-Golgi network transport, shaping downstream signaling and nutrient uptake. This pathway influences the homeostasis of receptor tyrosine kinases, GPCRs, and immune receptors, linking CLTC activity to proliferative and stress-response signaling networks. Genetic alterations and dysregulated clathrin-dependent trafficking have been associated with neurodevelopmental phenotypes and malignancy-related rearrangements involving CLTC, supporting mechanistic studies of vesicle transport defects in disease models.

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

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

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