transferrin CRISPR Activation Plasmid (h): sc-400871-ACT

Human TF encodes transferrin, a secreted glycoprotein that binds ferric iron in plasma and delivers it to cells through transferrin receptor–mediated endocytosis, supporting mitochondrial respiration, DNA synthesis, and cell-cycle progression. Transferrin-dependent iron trafficking integrates with iron homeostasis programs governed by IRP/IRE post-transcriptional regulation and interfaces with oxidative stress responses through iron-catalyzed redox chemistry. Dysregulated transferrin availability or TF expression is associated with altered systemic iron distribution, inflammatory signaling, and iron-loading or iron-restricted states that influence hematopoiesis and tissue metabolism. TF is therefore widely studied in nutrient sensing, ferroptosis-adjacent lipid peroxidation susceptibility, and tumor microenvironment iron dynamics in cell and molecular biology models.

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

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

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