Clathrin Light Chain B/CLTB CRISPR Activation Plasmid (h): sc-404228-ACT

CLTB encodes clathrin light chain B, a core component of clathrin-coated vesicles that regulates receptor-mediated endocytosis and intracellular trafficking between the plasma membrane, endosomes, and the trans-Golgi network. By modulating clathrin lattice dynamics with clathrin heavy chain and adaptor complexes such as AP2, CLTB contributes to cargo selection, vesicle budding, and synaptic vesicle recycling in specialized cell types. This trafficking axis influences signaling duration and receptor turnover, linking CLTB-associated processes to cellular homeostasis, proliferation, and stress responses. Dysregulated endocytic trafficking and clathrin-dependent transport are frequently studied in the context of neurobiology, viral entry pathways, and cancer-related receptor signaling alterations.

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

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

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