CSP CRISPR Activation Plasmid (h): sc-403622-ACT

Human DNAJC5 encodes cysteine string protein (CSP), a presynaptic co-chaperone that partners with HSC70 and other proteostasis factors to maintain synaptic vesicle cycling and neurotransmitter release. CSP supports the folding and stability of components of the SNARE/exocytosis machinery and helps safeguard nerve terminals from activity-dependent stress through chaperone-mediated quality control. By regulating vesicle trafficking and synaptic maintenance pathways, DNAJC5 is broadly relevant to studies of neuronal homeostasis and protein handling at the synapse. Altered CSP function is linked to neurodegenerative phenotypes characterized by synaptic dysfunction and disrupted protein turnover, making this gene useful for mechanistic models of neuronal vulnerability.

CSP CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous DNAJC5 expression without altering the underlying DNA sequence.

CSP CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the DNAJC5 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 DNAJC5 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous CSP expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native DNAJC5 locus and enabling the study of CSP-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of CSP pathway restoration in tumor cells with silenced or reduced DNAJC5 expression.

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