GTPBP4 CRISPR Activation Plasmid (h): sc-408845-ACT

Human GTPBP4 encodes a nucleolar P-loop NTPase that associates with pre-ribosomal particles and supports maturation of the 60S large ribosomal subunit. Through its roles in rRNA processing and ribosome biogenesis, GTPBP4 influences translational capacity, proteostasis, and cell-cycle progression under growth and stress conditions. Perturbation of nucleolar function and ribosome assembly is a hallmark of multiple proliferative and stress-response states, making GTPBP4 a useful node for dissecting links between nucleolar surveillance, p53-associated signaling, and global translation control. Aberrant regulation of ribosome biogenesis factors, including GTPases such as GTPBP4, has been connected in the literature to oncogenic programs and altered cellular fitness, supporting its relevance for mechanistic studies in disease models.

GTPBP4 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous GTPBP4 expression without altering the underlying DNA sequence.

GTPBP4 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the GTPBP4 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 GTPBP4 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous GTPBP4 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native GTPBP4 locus and enabling the study of GTPBP4-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of GTPBP4 pathway restoration in tumor cells with silenced or reduced GTPBP4 expression.

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