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Genomic stability of self-inactivating rabies
Transsynaptic viral vectors provide means to gain genetic access to neurons based on synaptic connectivity and are essential tools for the dissection of neural circuit function. Among them, the retrograde monosynaptic ΔG-Rabies has been widely used in neuroscience research. A recently developed engi...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
eLife Sciences Publications, Ltd
2023
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10666929/ https://www.ncbi.nlm.nih.gov/pubmed/37921437 http://dx.doi.org/10.7554/eLife.83459 |
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author | Ciabatti, Ernesto González-Rueda, Ana de Malmazet, Daniel Lee, Hassal Morgese, Fabio Tripodi, Marco |
author_facet | Ciabatti, Ernesto González-Rueda, Ana de Malmazet, Daniel Lee, Hassal Morgese, Fabio Tripodi, Marco |
author_sort | Ciabatti, Ernesto |
collection | PubMed |
description | Transsynaptic viral vectors provide means to gain genetic access to neurons based on synaptic connectivity and are essential tools for the dissection of neural circuit function. Among them, the retrograde monosynaptic ΔG-Rabies has been widely used in neuroscience research. A recently developed engineered version of the ΔG-Rabies, the non-toxic self-inactivating (SiR) virus, allows the long term genetic manipulation of neural circuits. However, the high mutational rate of the rabies virus poses a risk that mutations targeting the key genetic regulatory element in the SiR genome could emerge and revert it to a canonical ΔG-Rabies. Such revertant mutations have recently been identified in a SiR batch. To address the origin, incidence and relevance of these mutations, we investigated the genomic stability of SiR in vitro and in vivo. We found that “revertant” mutations are rare and accumulate only when SiR is extensively amplified in vitro, particularly in suboptimal production cell lines that have insufficient levels of TEV protease activity. Moreover, we confirmed that SiR-CRE, unlike canonical ΔG-Rab-CRE or revertant-SiR-CRE, is non-toxic and that revertant mutations do not emerge in vivo during long-term experiments. |
format | Online Article Text |
id | pubmed-10666929 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | eLife Sciences Publications, Ltd |
record_format | MEDLINE/PubMed |
spelling | pubmed-106669292023-11-03 Genomic stability of self-inactivating rabies Ciabatti, Ernesto González-Rueda, Ana de Malmazet, Daniel Lee, Hassal Morgese, Fabio Tripodi, Marco eLife Genetics and Genomics Transsynaptic viral vectors provide means to gain genetic access to neurons based on synaptic connectivity and are essential tools for the dissection of neural circuit function. Among them, the retrograde monosynaptic ΔG-Rabies has been widely used in neuroscience research. A recently developed engineered version of the ΔG-Rabies, the non-toxic self-inactivating (SiR) virus, allows the long term genetic manipulation of neural circuits. However, the high mutational rate of the rabies virus poses a risk that mutations targeting the key genetic regulatory element in the SiR genome could emerge and revert it to a canonical ΔG-Rabies. Such revertant mutations have recently been identified in a SiR batch. To address the origin, incidence and relevance of these mutations, we investigated the genomic stability of SiR in vitro and in vivo. We found that “revertant” mutations are rare and accumulate only when SiR is extensively amplified in vitro, particularly in suboptimal production cell lines that have insufficient levels of TEV protease activity. Moreover, we confirmed that SiR-CRE, unlike canonical ΔG-Rab-CRE or revertant-SiR-CRE, is non-toxic and that revertant mutations do not emerge in vivo during long-term experiments. eLife Sciences Publications, Ltd 2023-11-03 /pmc/articles/PMC10666929/ /pubmed/37921437 http://dx.doi.org/10.7554/eLife.83459 Text en © 2023, Ciabatti et al https://creativecommons.org/licenses/by/4.0/This article is distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use and redistribution provided that the original author and source are credited. |
spellingShingle | Genetics and Genomics Ciabatti, Ernesto González-Rueda, Ana de Malmazet, Daniel Lee, Hassal Morgese, Fabio Tripodi, Marco Genomic stability of self-inactivating rabies |
title | Genomic stability of self-inactivating rabies |
title_full | Genomic stability of self-inactivating rabies |
title_fullStr | Genomic stability of self-inactivating rabies |
title_full_unstemmed | Genomic stability of self-inactivating rabies |
title_short | Genomic stability of self-inactivating rabies |
title_sort | genomic stability of self-inactivating rabies |
topic | Genetics and Genomics |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10666929/ https://www.ncbi.nlm.nih.gov/pubmed/37921437 http://dx.doi.org/10.7554/eLife.83459 |
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