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RNAi and chemogenetic reporter co-regulation in primate striatal interneurons
Using genetic tools to study the functional roles of molecularly specified neuronal populations in the primate brain is challenging, primarily because of specificity and verification of virus-mediated targeting. Here, we report a lentivirus-based system that helps improve specificity and verificatio...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group UK
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8856958/ https://www.ncbi.nlm.nih.gov/pubmed/34012109 http://dx.doi.org/10.1038/s41434-021-00260-y |
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author | Lerchner, Walter Adil, Abdullah A. Mumuney, Sekinat Wang, Wenliang Falcone, Rossella Turchi, Janita Richmond, Barry J. |
author_facet | Lerchner, Walter Adil, Abdullah A. Mumuney, Sekinat Wang, Wenliang Falcone, Rossella Turchi, Janita Richmond, Barry J. |
author_sort | Lerchner, Walter |
collection | PubMed |
description | Using genetic tools to study the functional roles of molecularly specified neuronal populations in the primate brain is challenging, primarily because of specificity and verification of virus-mediated targeting. Here, we report a lentivirus-based system that helps improve specificity and verification by (a) targeting a selected molecular mechanism, (b) in vivo reporting of expression, and (c) allowing the option to independently silence all regional neural activity. Specifically, we modulate cholinergic signaling of striatal interneurons by shRNAmir and pair it with hM4Di_CFP, a chemogenetic receptor that can function as an in vivo and in situ reporter. Quantitative analyses by visual and deep-learning assisted methods show an inverse linear relation between hM4Di_CFP and ChAT protein expression for several shRNAmir constructs. This approach successfully applies shRNAmir to modulating gene expression in the primate brain and shows that hM4Di_CFP can act as a readout for this modulation. |
format | Online Article Text |
id | pubmed-8856958 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | Nature Publishing Group UK |
record_format | MEDLINE/PubMed |
spelling | pubmed-88569582022-03-10 RNAi and chemogenetic reporter co-regulation in primate striatal interneurons Lerchner, Walter Adil, Abdullah A. Mumuney, Sekinat Wang, Wenliang Falcone, Rossella Turchi, Janita Richmond, Barry J. Gene Ther Article Using genetic tools to study the functional roles of molecularly specified neuronal populations in the primate brain is challenging, primarily because of specificity and verification of virus-mediated targeting. Here, we report a lentivirus-based system that helps improve specificity and verification by (a) targeting a selected molecular mechanism, (b) in vivo reporting of expression, and (c) allowing the option to independently silence all regional neural activity. Specifically, we modulate cholinergic signaling of striatal interneurons by shRNAmir and pair it with hM4Di_CFP, a chemogenetic receptor that can function as an in vivo and in situ reporter. Quantitative analyses by visual and deep-learning assisted methods show an inverse linear relation between hM4Di_CFP and ChAT protein expression for several shRNAmir constructs. This approach successfully applies shRNAmir to modulating gene expression in the primate brain and shows that hM4Di_CFP can act as a readout for this modulation. Nature Publishing Group UK 2021-05-19 2022 /pmc/articles/PMC8856958/ /pubmed/34012109 http://dx.doi.org/10.1038/s41434-021-00260-y Text en © This is a U.S. government work and not under copyright protection in the U.S.; foreign copyright protection may apply 2021 https://creativecommons.org/licenses/by/4.0/Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as 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 images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . |
spellingShingle | Article Lerchner, Walter Adil, Abdullah A. Mumuney, Sekinat Wang, Wenliang Falcone, Rossella Turchi, Janita Richmond, Barry J. RNAi and chemogenetic reporter co-regulation in primate striatal interneurons |
title | RNAi and chemogenetic reporter co-regulation in primate striatal interneurons |
title_full | RNAi and chemogenetic reporter co-regulation in primate striatal interneurons |
title_fullStr | RNAi and chemogenetic reporter co-regulation in primate striatal interneurons |
title_full_unstemmed | RNAi and chemogenetic reporter co-regulation in primate striatal interneurons |
title_short | RNAi and chemogenetic reporter co-regulation in primate striatal interneurons |
title_sort | rnai and chemogenetic reporter co-regulation in primate striatal interneurons |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8856958/ https://www.ncbi.nlm.nih.gov/pubmed/34012109 http://dx.doi.org/10.1038/s41434-021-00260-y |
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