AFP/Alpha fetoprotein CRISPR Activation Plasmid (h): sc-400355-ACT

AFP encodes alpha-fetoprotein, a major oncofetal secreted glycoprotein produced predominantly by the fetal liver and yolk sac and downregulated after birth. AFP contributes to developmental programs by supporting hepatoblast differentiation and modulating extracellular signaling cues that influence proliferation and survival. In adult tissues, aberrant AFP re-expression is associated with hepatocellular lineage de-differentiation and altered transcriptional states observed in liver malignancy and related hepatic disorders. As a secreted biomarker-like protein, AFP is frequently used to study liver development, hepatic cell state transitions, and tumor-associated secretory phenotypes.

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

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

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