PIPELINE | CV-10,11
Terminally differentiated somatic cells can be reprogrammed to become induced pluripotent stem cells (iPSCs) by enforced expression of reprogramming factors (RFs) whichpromote self-renewal and render pluripotent cells leading to cellular differentiation. RFs contain 2 types of proteins, one for maintaining embryonic stem (ES) cells in a pluripotent state (OCT4, SOX2, and NANOG) and the other for promoting self-renewal and suppress cellular differentiation (CMYC, KLF4 and LIN28).
Based on this discovery, therapeutic approaches were developed by using autologous stem cells from patient-derived iPSCs without the ethical and graft rejection problems associated with using embryo-derived stem cells. Practically, the application of iPSCs to human regenerative medicine might require gene transfer to introduce RFs into somatic cells. Unfortunately, somatic cell reprogramming through gene transfer is relatively inefficient and is potentially mutagenic due to the genome integration of DNA-based expression vector. Other approaches that are not related toDNA-based expression vectors, such as, synthetic modified RNA and epigenetic regulation by chemical compounds are far from the practical application.
To overcome the limitation, we have newly invented improved cell-permeable RFs recombinant proteins (iCP-RFs) fused to novel hydrophobic CPP, advanced macromolecule transduction domain (aMTD), and solubilization domain (SD).
iCP-RFs induce stem cell-like colonies with high efficiency (0.01%~0.1%) and early colony formation with extension of self-renewal capacity and expression of stem cell-specific markers (OCT4, NANOG, TRA-1-60, TRA-1-81). Furthermore, by confirming that the stem cell-like colonies generated by iCP-RFs from stably differentiated teratoma of three germ layer, iCP-RFs have been shown to induce stem cells with complete pluripotent properties.
On the other hand, genome engineering is a technology for gene editing or repairing to recognize specific genetic sequence. This genome engineering technology is applicable to various fields, such as food, medical and research reagent. Especially, genome editing technology is used for therapeutic models to apply to therapy of genetic diseases.
The recent technology of CRISPR/Cas9 is powerful toolfor genome editing to repair disease-causing DNA mutations. However, a safe and efficient DNA delivery system are critical for guarantying the success of gene editing. Cell-permeable (CP) Cas9 with TSDT enables a simple treatment by gene editing with high cell-permeability. It is an innovative reagent that made it possible to be delivered into cells without DNA delivery. CP-Cas9 is increased in gene editing efficiency caused by intracellular transduction compared to previously used plasmid and mRNA system. These advantages of CP-Cas9 can be applied to gene modification as well as genetic research. CP-Cas9 also can be applied to stem cell therapy, which uses induced pluripotent stem cells (iPSC) for patients with genetic disorders.
Cas9 technology has been recently applied to disease-focused research through the production and characterization of patient-derived iPSCs from individuals with specific genetic diseases. The invention of induced iPSCs has greatly advanced translational research, especially with the generation of disease-derived human iPSCs. We believe CP-Cas9 will bring tremendous change forward in genetic transformation animal models, and even in stem cell research and therapy. Cellivery is open to out-licensing opportunities to institutions that want to begin research or development with CP-Cas9.