Transaldolase CRISPR Activation Plasmid (h): sc-402807-ACT

Human TALDO1 encodes transaldolase, a key enzyme of the non-oxidative branch of the pentose phosphate pathway that interconverts sugar phosphates to balance ribose-5-phosphate production with glycolytic intermediates. Through this role, TALDO1 supports nucleotide biosynthesis, redox homeostasis indirectly via pathway flux, and cellular adaptation to oxidative and metabolic stress. Altered transaldolase activity has been linked to inborn errors of metabolism and broader perturbations in hepatocellular function, immune regulation, and oxidative damage pathways. As a metabolic node, TALDO1 is frequently studied in contexts where pentose phosphate pathway rewiring affects proliferation, differentiation, and stress responses.

Transaldolase CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous TALDO1 expression without altering the underlying DNA sequence.

Transaldolase CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the TALDO1 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 TALDO1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Transaldolase expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native TALDO1 locus and enabling the study of Transaldolase-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Transaldolase pathway restoration in tumor cells with silenced or reduced TALDO1 expression.

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