Dynein HC CRISPR Activation Plasmid (h): sc-400919-ACT

DYNC1H1 encodes the cytoplasmic dynein 1 heavy chain, a microtubule minus-end–directed motor that powers long-range transport of vesicles, organelles, mRNAs, and protein complexes. Dynein heavy chain activity supports mitotic spindle organization, centrosome positioning, endosome/lysosome trafficking, and retrograde axonal transport through coordinated interactions with dynactin and cargo adaptors. As a core component of the cytoskeletal transport machinery, DYNC1H1 is integral to neuronal maintenance and intracellular logistics that shape polarity, proteostasis, and stress responses. Genetic perturbations of DYNC1H1 have been linked to neurodevelopmental and neuromuscular phenotypes, making it relevant for studies of axonopathy, motor neuron vulnerability, and transport-dependent signaling.

Dynein HC CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous DYNC1H1 expression without altering the underlying DNA sequence.

Dynein HC CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the DYNC1H1 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 DYNC1H1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Dynein HC expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native DYNC1H1 locus and enabling the study of Dynein HC-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Dynein HC pathway restoration in tumor cells with silenced or reduced DYNC1H1 expression.

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