Genomic stability of self-inactivating rabies

• DOI: 10.7554/elife.83459
• Publication Date: 2023-11-03T12:40:06Z
• Authors: Ernesto Ciabatti; Ana González-Rueda; Daniel de Malmazet; Hassal Lee; Fabio Morgese; Marco Tripodi
• Keywords: 0301 basic medicine; QH301-705.5; Rabies; Science; viral vector; neuronal tracing; Q; R; rabies; Genetics and Genomics; Genomic Instability; 3. Good health; Cell Line; neuroscience; 03 medical and health sciences; Rabies virus; Mutation; genomics; Medicine; Humans; genetics; Biology (General); mouse; neural circuits

Transsynaptic viral vectors provide means to gain genetic access neurons based on synaptic connectivity and are essential tools for the dissection of neural circuit function. Among them, retrograde monosynaptic ΔG-Rabies has been widely used in neuroscience research. A recently developed engineered version ΔG-Rabies, non-toxic self-inactivating (SiR) virus, allows long term manipulation circuits. However, high mutational rate rabies virus poses a risk that mutations targeting key regulatory element SiR genome could emerge revert it canonical ΔG-Rabies. Such revertant have identified batch. To address origin, incidence relevance these mutations, we investigated genomic stability vitro vivo. We found “revertant” rare accumulate only when is extensively amplified vitro, particularly suboptimal production cell lines insufficient levels TEV protease activity. Moreover, confirmed SiR-CRE, unlike ΔG-Rab-CRE or revertant-SiR-CRE, do not vivo during long-term experiments.