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High-brightness anterograde transneuronal HSV1 H129 tracer modified using a Trojan horse-like strategy

Neurotropic viral transsynaptic tracing is an increasingly powerful technique for dissecting the structure and function of neural circuits. Herpes simplex virus type 1 strain H129 has been widely used as an anterograde tracer. However, HSV tracers still have several shortcomings, including high toxi...

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Autores principales: Su, Peng, Ying, Min, Han, Zengpeng, Xia, Jinjin, Jin, Sen, Li, Yingli, Wang, Huadong, Xu, Fuqiang
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6958791/
https://www.ncbi.nlm.nih.gov/pubmed/31931837
http://dx.doi.org/10.1186/s13041-020-0544-2
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author Su, Peng
Ying, Min
Han, Zengpeng
Xia, Jinjin
Jin, Sen
Li, Yingli
Wang, Huadong
Xu, Fuqiang
author_facet Su, Peng
Ying, Min
Han, Zengpeng
Xia, Jinjin
Jin, Sen
Li, Yingli
Wang, Huadong
Xu, Fuqiang
author_sort Su, Peng
collection PubMed
description Neurotropic viral transsynaptic tracing is an increasingly powerful technique for dissecting the structure and function of neural circuits. Herpes simplex virus type 1 strain H129 has been widely used as an anterograde tracer. However, HSV tracers still have several shortcomings, including high toxicity, low sensitivity and non-specific retrograde labeling. Here, we aimed to construct high-brightness HSV anterograde tracers by increasing the expression of exogenous genes carried by H129 viruses. Using a Trojan horse-like strategy, a HSV/AAV (adeno-associated virus) chimaera termed H8 was generated to enhance the expression of a fluorescent marker. In vitro and in vivo assays showed that the exogenous gene was efficiently replicated and amplified by the synergism of the HSV vector and introduced AAV replication system. H8 reporting fluorescence was brighter than that of currently available H129 tracers, and H8 could be used for fast and effective anterograde tracing without additional immunostaining. These results indicated that foreign gene expression in HSV tracers could be enhanced by integrating HSV with AAV replication system. This approach may be useful as a general enhanced expression strategy for HSV-based tracing tools or gene delivery vectors.
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spelling pubmed-69587912020-01-17 High-brightness anterograde transneuronal HSV1 H129 tracer modified using a Trojan horse-like strategy Su, Peng Ying, Min Han, Zengpeng Xia, Jinjin Jin, Sen Li, Yingli Wang, Huadong Xu, Fuqiang Mol Brain Methodology Neurotropic viral transsynaptic tracing is an increasingly powerful technique for dissecting the structure and function of neural circuits. Herpes simplex virus type 1 strain H129 has been widely used as an anterograde tracer. However, HSV tracers still have several shortcomings, including high toxicity, low sensitivity and non-specific retrograde labeling. Here, we aimed to construct high-brightness HSV anterograde tracers by increasing the expression of exogenous genes carried by H129 viruses. Using a Trojan horse-like strategy, a HSV/AAV (adeno-associated virus) chimaera termed H8 was generated to enhance the expression of a fluorescent marker. In vitro and in vivo assays showed that the exogenous gene was efficiently replicated and amplified by the synergism of the HSV vector and introduced AAV replication system. H8 reporting fluorescence was brighter than that of currently available H129 tracers, and H8 could be used for fast and effective anterograde tracing without additional immunostaining. These results indicated that foreign gene expression in HSV tracers could be enhanced by integrating HSV with AAV replication system. This approach may be useful as a general enhanced expression strategy for HSV-based tracing tools or gene delivery vectors. BioMed Central 2020-01-13 /pmc/articles/PMC6958791/ /pubmed/31931837 http://dx.doi.org/10.1186/s13041-020-0544-2 Text en © The Author(s). 2020 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Methodology
Su, Peng
Ying, Min
Han, Zengpeng
Xia, Jinjin
Jin, Sen
Li, Yingli
Wang, Huadong
Xu, Fuqiang
High-brightness anterograde transneuronal HSV1 H129 tracer modified using a Trojan horse-like strategy
title High-brightness anterograde transneuronal HSV1 H129 tracer modified using a Trojan horse-like strategy
title_full High-brightness anterograde transneuronal HSV1 H129 tracer modified using a Trojan horse-like strategy
title_fullStr High-brightness anterograde transneuronal HSV1 H129 tracer modified using a Trojan horse-like strategy
title_full_unstemmed High-brightness anterograde transneuronal HSV1 H129 tracer modified using a Trojan horse-like strategy
title_short High-brightness anterograde transneuronal HSV1 H129 tracer modified using a Trojan horse-like strategy
title_sort high-brightness anterograde transneuronal hsv1 h129 tracer modified using a trojan horse-like strategy
topic Methodology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6958791/
https://www.ncbi.nlm.nih.gov/pubmed/31931837
http://dx.doi.org/10.1186/s13041-020-0544-2
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