NTF2 CRISPR Activation Plasmid (h): sc-405048-ACT

Human NUTF2 encodes nuclear transport factor 2 (NTF2), a key mediator of nucleocytoplasmic transport that supports nuclear import by regulating the Ran GTPase cycle and facilitating translocation of RanGDP through the nuclear pore complex. By maintaining the RanGTP/RanGDP gradient, NTF2 influences broad processes including cell-cycle progression, stress responses, and the nuclear localization of regulatory proteins and RNAs. Altered nuclear transport dynamics and Ran pathway imbalance are frequently observed in proliferative and neurodegenerative disease contexts, making NUTF2 a useful node for dissecting transport-dependent signaling and gene regulation. NTF2 function is also relevant to studies of nuclear pore biology and host–pathogen interactions that exploit nuclear trafficking.

NTF2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous NUTF2 expression without altering the underlying DNA sequence.

NTF2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the NUTF2 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 NUTF2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous NTF2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native NUTF2 locus and enabling the study of NTF2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of NTF2 pathway restoration in tumor cells with silenced or reduced NUTF2 expression.

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