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Leaky expression of channelrhodopsin-2 (ChR2) in Ai32 mouse lines
Optogenetics enables the selective activation of genetically-targeted neuronal populations using light-sensitive ion channels. Genetic strategies using Cre-dependent mouse strains, especially the Ai32 line expressing Channelrhodopsin (ChR2)-EYFP fusion protein, have been a popular means to drive ops...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2019
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6435231/ https://www.ncbi.nlm.nih.gov/pubmed/30913225 http://dx.doi.org/10.1371/journal.pone.0213326 |
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author | Prabhakar, Arthi Vujovic, Dragan Cui, Lian Olson, William Luo, Wenqin |
author_facet | Prabhakar, Arthi Vujovic, Dragan Cui, Lian Olson, William Luo, Wenqin |
author_sort | Prabhakar, Arthi |
collection | PubMed |
description | Optogenetics enables the selective activation of genetically-targeted neuronal populations using light-sensitive ion channels. Genetic strategies using Cre-dependent mouse strains, especially the Ai32 line expressing Channelrhodopsin (ChR2)-EYFP fusion protein, have been a popular means to drive opsin expression in a cell-type specific manner. Here we report a low level of leaky ‘off-target’ (Cre-independent) ChR2-EYFP expression in Ai32/Ai32 homozygous mice throughout the nervous system. This leaky off-target expression was characterized in multiple prevalent nervous system regions using anti-EYFP immunostaining. Expression of full-length ChR2-EYFP protein was confirmed using immunoprecipitation followed by Western blotting. Notably, light stimulation of these ChR2-EYFP expressing neurons in the spinal cord dorsal horn did not induce detectable photocurrents in juvenile 4-week old mice. Given the wide use of the Ai32 line by many labs, our results suggest researchers should be vigilant of possible off-target ChR2-EYFP expression in their region of interest, especially when generating Ai32/Ai32 homozygotes to drive high levels of ChR2-EYFP expression in adult mice. |
format | Online Article Text |
id | pubmed-6435231 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-64352312019-04-08 Leaky expression of channelrhodopsin-2 (ChR2) in Ai32 mouse lines Prabhakar, Arthi Vujovic, Dragan Cui, Lian Olson, William Luo, Wenqin PLoS One Research Article Optogenetics enables the selective activation of genetically-targeted neuronal populations using light-sensitive ion channels. Genetic strategies using Cre-dependent mouse strains, especially the Ai32 line expressing Channelrhodopsin (ChR2)-EYFP fusion protein, have been a popular means to drive opsin expression in a cell-type specific manner. Here we report a low level of leaky ‘off-target’ (Cre-independent) ChR2-EYFP expression in Ai32/Ai32 homozygous mice throughout the nervous system. This leaky off-target expression was characterized in multiple prevalent nervous system regions using anti-EYFP immunostaining. Expression of full-length ChR2-EYFP protein was confirmed using immunoprecipitation followed by Western blotting. Notably, light stimulation of these ChR2-EYFP expressing neurons in the spinal cord dorsal horn did not induce detectable photocurrents in juvenile 4-week old mice. Given the wide use of the Ai32 line by many labs, our results suggest researchers should be vigilant of possible off-target ChR2-EYFP expression in their region of interest, especially when generating Ai32/Ai32 homozygotes to drive high levels of ChR2-EYFP expression in adult mice. Public Library of Science 2019-03-26 /pmc/articles/PMC6435231/ /pubmed/30913225 http://dx.doi.org/10.1371/journal.pone.0213326 Text en © 2019 Prabhakar et al http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. |
spellingShingle | Research Article Prabhakar, Arthi Vujovic, Dragan Cui, Lian Olson, William Luo, Wenqin Leaky expression of channelrhodopsin-2 (ChR2) in Ai32 mouse lines |
title | Leaky expression of channelrhodopsin-2 (ChR2) in Ai32 mouse lines |
title_full | Leaky expression of channelrhodopsin-2 (ChR2) in Ai32 mouse lines |
title_fullStr | Leaky expression of channelrhodopsin-2 (ChR2) in Ai32 mouse lines |
title_full_unstemmed | Leaky expression of channelrhodopsin-2 (ChR2) in Ai32 mouse lines |
title_short | Leaky expression of channelrhodopsin-2 (ChR2) in Ai32 mouse lines |
title_sort | leaky expression of channelrhodopsin-2 (chr2) in ai32 mouse lines |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6435231/ https://www.ncbi.nlm.nih.gov/pubmed/30913225 http://dx.doi.org/10.1371/journal.pone.0213326 |
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