TBC1D24 CRISPR Activation Plasmid (h): sc-404561-ACT

TBC1D24 encodes a TBC domain–containing protein implicated in regulation of membrane trafficking and vesicle dynamics, processes that support synaptic function and other high-demand secretory pathways. Although its TBC domain is atypical compared with canonical Rab GTPase-activating proteins, TBC1D24 has been linked to control of endosomal sorting, synaptic vesicle cycling, and maintenance of cellular homeostasis under oxidative stress. In neurons, these pathways intersect with activity-dependent signaling and organelle turnover that influence excitability and network stability. Genetic variation in TBC1D24 is associated with neurodevelopmental and epileptic phenotypes, motivating mechanistic studies of dosage- and context-dependent effects in relevant cellular models.

TBC1D24 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous TBC1D24 expression without altering the underlying DNA sequence.

TBC1D24 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the TBC1D24 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 TBC1D24 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous TBC1D24 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native TBC1D24 locus and enabling the study of TBC1D24-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of TBC1D24 pathway restoration in tumor cells with silenced or reduced TBC1D24 expression.

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