TBC1D7 CRISPR Activation Plasmid (h): sc-403595-ACT

TBC1D7 encodes a small TBC domain–containing protein that functions as a core component of the TSC1–TSC2–TBC1D7 complex, a key negative regulator of mTORC1 signaling. By stabilizing the TSC complex and supporting GAP activity toward RHEB, TBC1D7 helps couple growth factor and nutrient cues to control of protein synthesis, autophagy, and cellular metabolism. Altered TBC1D7 dosage or function has been linked to dysregulated mTOR pathway activity and phenotypes consistent with tumor suppressor network impairment. As a result, TBC1D7 is widely studied in contexts such as cell growth control, stress responses, and pathway crosstalk influencing proliferation and differentiation.

TBC1D7 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous TBC1D7 expression without altering the underlying DNA sequence.

TBC1D7 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the TBC1D7 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 TBC1D7 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous TBC1D7 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native TBC1D7 locus and enabling the study of TBC1D7-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of TBC1D7 pathway restoration in tumor cells with silenced or reduced TBC1D7 expression.

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