CD71/TFRC/Transferrin Receptor CRISPR Activation Plasmid (h): sc-400310-ACT

TFRC encodes CD71, the transferrin receptor that mediates high-affinity binding and clathrin-dependent endocytosis of transferrin to deliver iron to cells, supporting heme synthesis, mitochondrial respiration, and DNA replication. Following endosomal acidification and iron release, apo-transferrin and CD71 recycle to the plasma membrane, coupling iron uptake to membrane trafficking and lysosomal/endosomal pathways. TFRC expression is tightly regulated by iron-responsive elements and integrates with IRP/IRE signaling, MYC-driven proliferation programs, and hypoxia-responsive networks that tune cellular iron demand. Dysregulated TFRC activity and iron handling are frequently studied in cancer metabolism, immune cell activation, anemia-related biology, and host–pathogen interactions that depend on iron availability.

CD71/TFRC/Transferrin Receptor CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous TFRC expression without altering the underlying DNA sequence.

CD71/TFRC/Transferrin Receptor CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the TFRC 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 TFRC transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous CD71/TFRC/Transferrin Receptor expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native TFRC locus and enabling the study of CD71/TFRC/Transferrin Receptor-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of CD71/TFRC/Transferrin Receptor pathway restoration in tumor cells with silenced or reduced TFRC expression.

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