Hemophilia B can be treated by iPSC differentiated hepatocytes with AAVS1 safe harborknockin Coagulation factor IX (F9)
The most common method to treat hemophilia is substitution therapy, but this method has the risk of virus infection, and it is a method that needs lifelong continuous treatment. Gene therapy seems like the only way can cure hemophilia. CRISPR/Cas9 technology can be used for gene therapy of hemophilia. The mutations of coagulation factors, F8 and F9, are the main causes of hemophilia. Previous studies have shown that F9 is a more effective gene therapy target. AAVS1-Cas9-sgRNAplasmid and AAVS1-EF1α-F9 cDNA puromycin donor plasmid were constructed and transferred into iPSC. Human factor IX (hFIX) antigen activity was detected in the culture supernatant. Finally, liver cells differentiated from iPSC were transplanted into NOD/SCIDmice by spleen injection, to cure hemophilia B.（Lyu, Cuicui, et al.）
With CRISPR-UTM, iPSCwould be co-transfected with gRNA, Cas9 and donor vector by electroporation. After drug screening, single clones would be generated. Positive clones would be validated by sequencing.
iPSC induced differentiation, making "autotransplantation" possible
The study of human embryonic stem cells (hESCs) derived from early embryos has been controversial in ethics, and the rejection of differentiated cells derived from hESCs intransplantation has limited its clinical application. Hepatocytes, nerve cells, T cells, cardiomyocytes, hematopoietic stem cells and pancreatic cells can be differentiated from patients' somatic cells (such as fibroblasts) or existing iPSCs.
iPSC Differentiation Process
iPSC reprogramming: Ubigenehas optimized the reprogramming method ——>iPSC induced differentiation: continuous addition of inducing factor in iPSC medium——>Cell validation: genotype, phenotype and cell characteristic analysis
Ubigene's iPSC platform
Ubigene focuses on the optimization of iPSC reprogramming, gene editing and differentiation, and has established a set of mature experimental procedures. With CRIPSR-UTM technology, gene editing in iPSC is much accessible.