Cre-assisted fine-mapping of neural circuits using orthogonal split inteins
Existing genetic methods of neuronal targeting do not routinely achieve the resolution required for mapping brain circuits. New approaches are thus necessary. Here, we introduce a method for refined neuronal targeting that can be applied iteratively. Restriction achieved at the first step can be fur...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
| Published: |
eLife Sciences Publications Ltd
2020-04-01
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| Series: | eLife |
| Subjects: | |
| Online Access: | https://elifesciences.org/articles/53041 |
| _version_ | 1828221588918501376 |
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| author | Haojiang Luan Alexander Kuzin Ward F Odenwald Benjamin H White |
| author_facet | Haojiang Luan Alexander Kuzin Ward F Odenwald Benjamin H White |
| author_sort | Haojiang Luan |
| collection | DOAJ |
| description | Existing genetic methods of neuronal targeting do not routinely achieve the resolution required for mapping brain circuits. New approaches are thus necessary. Here, we introduce a method for refined neuronal targeting that can be applied iteratively. Restriction achieved at the first step can be further refined in a second step, if necessary. The method relies on first isolating neurons within a targeted group (i.e. Gal4 pattern) according to their developmental lineages, and then intersectionally limiting the number of lineages by selecting only those in which two distinct neuroblast enhancers are active. The neuroblast enhancers drive expression of split Cre recombinase fragments. These are fused to non-interacting pairs of split inteins, which ensure reconstitution of active Cre when all fragments are expressed in the same neuroblast. Active Cre renders all neuroblast-derived cells in a lineage permissive for Gal4 activity. We demonstrate how this system can facilitate neural circuit-mapping in Drosophila. |
| first_indexed | 2024-04-12T16:43:40Z |
| format | Article |
| id | doaj.art-cb871df6b4eb4acd86e2175bfb4f466a |
| institution | Directory Open Access Journal |
| issn | 2050-084X |
| language | English |
| last_indexed | 2024-04-12T16:43:40Z |
| publishDate | 2020-04-01 |
| publisher | eLife Sciences Publications Ltd |
| record_format | Article |
| series | eLife |
| spelling | doaj.art-cb871df6b4eb4acd86e2175bfb4f466a2022-12-22T03:24:41ZengeLife Sciences Publications LtdeLife2050-084X2020-04-01910.7554/eLife.53041Cre-assisted fine-mapping of neural circuits using orthogonal split inteinsHaojiang Luan0Alexander Kuzin1Ward F Odenwald2Benjamin H White3https://orcid.org/0000-0003-0612-8075Laboratory of Molecular Biology, National Institute of Mental Health, NIH, Bethesda, United StatesNeural Cell-Fate Determinants Section, National Institute of Neurological Disorders and Stroke, NIH, Bethesda, United StatesNeural Cell-Fate Determinants Section, National Institute of Neurological Disorders and Stroke, NIH, Bethesda, United StatesLaboratory of Molecular Biology, National Institute of Mental Health, NIH, Bethesda, United StatesExisting genetic methods of neuronal targeting do not routinely achieve the resolution required for mapping brain circuits. New approaches are thus necessary. Here, we introduce a method for refined neuronal targeting that can be applied iteratively. Restriction achieved at the first step can be further refined in a second step, if necessary. The method relies on first isolating neurons within a targeted group (i.e. Gal4 pattern) according to their developmental lineages, and then intersectionally limiting the number of lineages by selecting only those in which two distinct neuroblast enhancers are active. The neuroblast enhancers drive expression of split Cre recombinase fragments. These are fused to non-interacting pairs of split inteins, which ensure reconstitution of active Cre when all fragments are expressed in the same neuroblast. Active Cre renders all neuroblast-derived cells in a lineage permissive for Gal4 activity. We demonstrate how this system can facilitate neural circuit-mapping in Drosophila.https://elifesciences.org/articles/53041circuit mappingneuroblastGal4-UASproboscis extensiondopamineexpression system |
| spellingShingle | Haojiang Luan Alexander Kuzin Ward F Odenwald Benjamin H White Cre-assisted fine-mapping of neural circuits using orthogonal split inteins eLife circuit mapping neuroblast Gal4-UAS proboscis extension dopamine expression system |
| title | Cre-assisted fine-mapping of neural circuits using orthogonal split inteins |
| title_full | Cre-assisted fine-mapping of neural circuits using orthogonal split inteins |
| title_fullStr | Cre-assisted fine-mapping of neural circuits using orthogonal split inteins |
| title_full_unstemmed | Cre-assisted fine-mapping of neural circuits using orthogonal split inteins |
| title_short | Cre-assisted fine-mapping of neural circuits using orthogonal split inteins |
| title_sort | cre assisted fine mapping of neural circuits using orthogonal split inteins |
| topic | circuit mapping neuroblast Gal4-UAS proboscis extension dopamine expression system |
| url | https://elifesciences.org/articles/53041 |
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