TAP CRISPR Activation Plasmid (h): sc-402586-ACT

Human NXF1 encodes TAP, a central mRNA export receptor that couples transcription and pre-mRNA processing to nuclear pore transport by forming a heterodimer with NXT1 and engaging nucleoporins through FG-repeat interactions. TAP binds adaptor complexes such as TREX and recognizes mRNP features to promote efficient nuclear export, thereby influencing global gene expression, RNA surveillance, and coupling of splicing to export. Through its role in regulating the composition and cytoplasmic availability of mRNAs, NXF1/TAP impacts pathways including RNA metabolism, translation control, and stress-responsive gene expression programs. Dysregulation of mRNA export machinery has been associated with altered transcriptome homeostasis observed in cancer and neurodegenerative disease contexts, making NXF1 a relevant target for mechanistic studies of RNA transport and gene expression fidelity.

TAP CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous NXF1 expression without altering the underlying DNA sequence.

TAP CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the NXF1 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 NXF1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous TAP expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native NXF1 locus and enabling the study of TAP-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of TAP pathway restoration in tumor cells with silenced or reduced NXF1 expression.

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