KLC1 CRISPR Activation Plasmid (h): sc-401881-ACT

Human KLC1 (kinesin light chain 1) encodes a light chain subunit of the kinesin-1 motor complex that links cargo adaptors to microtubule-based transport. By partnering with kinesin heavy chains and cargo-binding proteins, KLC1 supports anterograde trafficking of vesicles, organelles, and protein complexes that is essential for neuronal polarity, axonal transport, and spatial control of signaling. KLC1-dependent transport interfaces with pathways governing endosomal trafficking, mitochondrial distribution, and synaptic function, where disrupted cargo delivery can perturb proteostasis and stress responses. Altered KLC1 expression or cargo coupling has been investigated in neurodegeneration and other disorders featuring transport defects and dysregulated intracellular trafficking.

KLC1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous KLC1 expression without altering the underlying DNA sequence.

KLC1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the KLC1 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 KLC1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous KLC1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native KLC1 locus and enabling the study of KLC1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of KLC1 pathway restoration in tumor cells with silenced or reduced KLC1 expression.

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