TPR CRISPR Activation Plasmid (h): sc-402928-ACT

Human TPR encodes a large coiled-coil nucleoporin that forms a key scaffold of the nuclear pore basket, supporting nuclear–cytoplasmic transport and higher-order nuclear organization. TPR contributes to mRNA export and surveillance, interfaces with the spindle checkpoint during mitosis, and helps maintain genome stability through coordination of nuclear envelope–associated processes. Through these roles, altered TPR regulation or fusion events have been linked to dysregulated signaling and chromosomal rearrangements observed in multiple cancer contexts, making it relevant to studies of oncogenic transcriptional programs and cell-cycle control. TPR function is also commonly examined in pathways governing RNA processing, mitotic fidelity, and stress-responsive nuclear trafficking.

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

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

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