CRISPR/Cas9-Based Edition of Frataxin Gene inDictyostelium discoideumfor Friedreich’s Ataxia Disease Modeling

Abstract

AbstractIn this paper we describe the development of a new model system for Friedreich’s Atax- ia (FA) usingDictyostelium discoideum. We investigated the conservation of function between humans andD. discoideumand showed that DdFXN can substitute the human version in the interaction and activation of the Fe-S assembly supercomplex. We edited thefxnlocus and isolated a defective mutant, clone 8, which presents landmarks of frataxin deficiency, such as a decrease in Fe-S cluster-dependent enzymatic functions, growth rate reduction, and increased sensitivity to oxidative stress. In addition multicellular development is affected as well as grow on bacterial lawn.We also assessed the rescuing capacity of DdFXN-G122V, a version that mimics a human variant present in some FA patients. While the expression of DdFXN-G122V rescues growth and enzymatic activity defects, as DdFXN does, multicellular development defects were only partially rescuedThe results of the study suggest that this new model system offers a wide range of pos- sibilities to easily explore diverse phenotypes in FA and develop drug or treatment screenings for designing and evaluating therapeutic strategies.