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“Self-inactivating” rabies viruses are susceptible to loss of their intended attenuating modification

Monosynaptic tracing using rabies virus is an important technique in neuroscience, allowing brain-wide labeling of neurons directly presynaptic to a targeted neuronal population. A 2017 article reported the development of a noncytotoxic version—a major advance—based on attenuating the rabies virus b...

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Autores principales: Jin, Lei, Matsuyama, Makoto, Sullivan, Heather A., Zhu, Mulangma, Lavin, Thomas K., Hou, YuanYuan, Lea, Nicholas E., Pruner, Maxwell T., Dam Ferdínez, María Lucía, Wickersham, Ian R.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: National Academy of Sciences 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9963760/
https://www.ncbi.nlm.nih.gov/pubmed/37053554
http://dx.doi.org/10.1073/pnas.2023481120
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author Jin, Lei
Matsuyama, Makoto
Sullivan, Heather A.
Zhu, Mulangma
Lavin, Thomas K.
Hou, YuanYuan
Lea, Nicholas E.
Pruner, Maxwell T.
Dam Ferdínez, María Lucía
Wickersham, Ian R.
author_facet Jin, Lei
Matsuyama, Makoto
Sullivan, Heather A.
Zhu, Mulangma
Lavin, Thomas K.
Hou, YuanYuan
Lea, Nicholas E.
Pruner, Maxwell T.
Dam Ferdínez, María Lucía
Wickersham, Ian R.
author_sort Jin, Lei
collection PubMed
description Monosynaptic tracing using rabies virus is an important technique in neuroscience, allowing brain-wide labeling of neurons directly presynaptic to a targeted neuronal population. A 2017 article reported the development of a noncytotoxic version—a major advance—based on attenuating the rabies virus by the addition of a destabilization domain to the C terminus of a viral protein. However, this modification did not appear to hinder the ability of the virus to spread between neurons. We analyzed two viruses provided by the authors and show here that both were mutants that had lost the intended modification, explaining the paper's paradoxical results. We then made a virus that actually did have the intended modification in at least the majority of virions and found that it did not spread efficiently under the conditions described in the original paper, namely, without an exogenous protease being expressed in order to remove the destabilization domain. We found that it did spread when the protease was supplied, although this also appeared to result in the deaths of most source cells by 3 wk postinjection. We conclude that the new approach is not robust but that it could become a viable technique given further optimization and validation.
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spelling pubmed-99637602023-08-06 “Self-inactivating” rabies viruses are susceptible to loss of their intended attenuating modification Jin, Lei Matsuyama, Makoto Sullivan, Heather A. Zhu, Mulangma Lavin, Thomas K. Hou, YuanYuan Lea, Nicholas E. Pruner, Maxwell T. Dam Ferdínez, María Lucía Wickersham, Ian R. Proc Natl Acad Sci U S A Biological Sciences Monosynaptic tracing using rabies virus is an important technique in neuroscience, allowing brain-wide labeling of neurons directly presynaptic to a targeted neuronal population. A 2017 article reported the development of a noncytotoxic version—a major advance—based on attenuating the rabies virus by the addition of a destabilization domain to the C terminus of a viral protein. However, this modification did not appear to hinder the ability of the virus to spread between neurons. We analyzed two viruses provided by the authors and show here that both were mutants that had lost the intended modification, explaining the paper's paradoxical results. We then made a virus that actually did have the intended modification in at least the majority of virions and found that it did not spread efficiently under the conditions described in the original paper, namely, without an exogenous protease being expressed in order to remove the destabilization domain. We found that it did spread when the protease was supplied, although this also appeared to result in the deaths of most source cells by 3 wk postinjection. We conclude that the new approach is not robust but that it could become a viable technique given further optimization and validation. National Academy of Sciences 2023-02-06 2023-02-14 /pmc/articles/PMC9963760/ /pubmed/37053554 http://dx.doi.org/10.1073/pnas.2023481120 Text en Copyright © 2023 the Author(s). Published by PNAS. https://creativecommons.org/licenses/by-nc-nd/4.0/This article is distributed under Creative Commons Attribution-NonCommercial-NoDerivatives License 4.0 (CC BY-NC-ND) (https://creativecommons.org/licenses/by-nc-nd/4.0/) .
spellingShingle Biological Sciences
Jin, Lei
Matsuyama, Makoto
Sullivan, Heather A.
Zhu, Mulangma
Lavin, Thomas K.
Hou, YuanYuan
Lea, Nicholas E.
Pruner, Maxwell T.
Dam Ferdínez, María Lucía
Wickersham, Ian R.
“Self-inactivating” rabies viruses are susceptible to loss of their intended attenuating modification
title “Self-inactivating” rabies viruses are susceptible to loss of their intended attenuating modification
title_full “Self-inactivating” rabies viruses are susceptible to loss of their intended attenuating modification
title_fullStr “Self-inactivating” rabies viruses are susceptible to loss of their intended attenuating modification
title_full_unstemmed “Self-inactivating” rabies viruses are susceptible to loss of their intended attenuating modification
title_short “Self-inactivating” rabies viruses are susceptible to loss of their intended attenuating modification
title_sort “self-inactivating” rabies viruses are susceptible to loss of their intended attenuating modification
topic Biological Sciences
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9963760/
https://www.ncbi.nlm.nih.gov/pubmed/37053554
http://dx.doi.org/10.1073/pnas.2023481120
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