RNGTT CRISPR Activation Plasmid (h): sc-405280-ACT

Human RNGTT encodes the RNA guanylyltransferase and 5′-phosphatase that catalyzes key steps of mRNA 5′ capping, converting nascent RNA 5′ triphosphates to the m7G cap in coordination with methyltransferases and the transcription machinery. This cap structure is essential for pre-mRNA processing, nuclear export, translation initiation, and protection from exonucleases, linking RNGTT activity to RNA polymerase II–coupled transcription and global gene expression programs. By influencing mRNA stability and translation efficiency, RNGTT intersects with pathways that govern cell-cycle progression, stress responses, and innate immune sensing of uncapped RNA. Dysregulated mRNA capping and cap-dependent translation control are broadly relevant to proliferative and neurodevelopmental disease biology, making RNGTT a useful node for mechanistic studies of RNA metabolism.

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

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

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