Â鶹Éç

Our understanding of genes and mutations critical for neuronal development and disease has vast knowledge gaps due to limitations of current research methods. We will therefore develop CRISPRNeuroFate, a disruptive technology that can identify such genes and mutations rapidly and conclusively in relevant neuronal model systems. 

Specifically, CRISPRNeuroFate simultaneously engineers a human pluripotent stem cell population with test mutations of interest (a STOP mutation and (optionally) a disease mutation) and two synonymous (neutral) control mutations (WT´) in gene of interest. Next, the stem cell population is differentiated into an organ/tissue of interest in vitro, in which CRISPRNeuroFate quantitatively tracks nerve cells with test mutations relative to nerve cells with control mutations in the developing organ as a function of time and expression of FACS markers of interest (cell type, differentiation, function, metabolism, apoptosis etc).  

This will conclusively determine how the test mutations impact the nerve cells of interest. As PoC, we will apply CRISPRNeuroFate to the human eye and identify genes essential for development of the optic nerve and neuronal photoreceptor cells as well as novel disease mutations causing their degeneration in the major eye diseases glaucoma and retinitis pigmentosa. Our approach may be expanded to any stem cell-based neuronal differentiation models and provide a game-changer technology for the study of development, function and genetic disease in the human nervous system.

The applicant Morten Frödin has developed several of the most widely used CRISPR techniques, including CRISPR editing of stem cells. Assistant prof. Anne Z. Eriksen is collaborator, an expert in stem cell biology with extensive experience in the eye development model in which Morten Frödin will develop the present technology as PoC. Anne Z. Eriksen just received the prestigious Sapere Aude research leader grant to study retinitis pigmentosa in the eye model.