Long-term precision editing of neural circuits in mammals using engineered gap junction hemichannels
Summary The coordination of activity between brain cells is a key determinant of neural circuit function; nevertheless, approaches that selectively regulate communication between two distinct cellular components of a mammalian circuit remain sparse. To address this gap, we developed a novel class of gap junctions by selectively engineering two connexin proteins found inMorone americana(white perch fish): connexin34.7 (Cx34.7) and connexin35 (Cx35). By iteratively exploiting protein mutagenesis, a novelin vitroassay of connexin docking, and computational modeling of connexin hemichannel interactions, we uncovered a pattern of structural motifs that contribute to hemichannel docking compatibility. Targeting these motifs, we designed Cx34.7 and Cx35 hemichannels that dock with each other, but not with themselves, nor with other major connexins expressed in the mammalian central nervous system. We validated these hemichannelsin vivousingC. elegansand mice, demonstrating that they can facilitate communication across neural circuits composed of pairs of distinct cell types and modify behavior accordingly. Thus, we establish a potentially translational approach, ‘<jats:underline>L</jats:underline>ong-term<jats:underline>in</jats:underline>tegration of<jats:underline>C</jats:underline>ircuits using conne<jats:underline>x</jats:underline>ins’ (LinCx), for context-precise circuit-editing with unprecedented spatiotemporal specificity in mammals..
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Preprint| Erscheinungsjahr: |
2023 |
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| Erschienen: |
2023 |
| Enthalten in: |
bioRxiv.org - (2023) vom: 22. März Zur Gesamtaufnahme - year:2023 |
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| Sprache: |
Englisch |
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| Beteiligte Personen: |
Ransey, Elizabeth [VerfasserIn] |
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| Links: |
Volltext [kostenfrei] |
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| doi: |
10.1101/2021.08.24.457429 |
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| PPN (Katalog-ID): |
XBI032467176 |
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| 245 | 1 | 0 | |a Long-term precision editing of neural circuits in mammals using engineered gap junction hemichannels |
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| 520 | |a Summary The coordination of activity between brain cells is a key determinant of neural circuit function; nevertheless, approaches that selectively regulate communication between two distinct cellular components of a mammalian circuit remain sparse. To address this gap, we developed a novel class of gap junctions by selectively engineering two connexin proteins found inMorone americana(white perch fish): connexin34.7 (Cx34.7) and connexin35 (Cx35). By iteratively exploiting protein mutagenesis, a novelin vitroassay of connexin docking, and computational modeling of connexin hemichannel interactions, we uncovered a pattern of structural motifs that contribute to hemichannel docking compatibility. Targeting these motifs, we designed Cx34.7 and Cx35 hemichannels that dock with each other, but not with themselves, nor with other major connexins expressed in the mammalian central nervous system. We validated these hemichannelsin vivousingC. elegansand mice, demonstrating that they can facilitate communication across neural circuits composed of pairs of distinct cell types and modify behavior accordingly. Thus, we establish a potentially translational approach, ‘<jats:underline>L</jats:underline>ong-term<jats:underline>in</jats:underline>tegration of<jats:underline>C</jats:underline>ircuits using conne<jats:underline>x</jats:underline>ins’ (LinCx), for context-precise circuit-editing with unprecedented spatiotemporal specificity in mammals. | ||
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| 700 | 1 | |a Chesnov, Kirill |4 aut | |
| 700 | 1 | |a Thomas, Gwenaëlle E. |4 aut | |
| 700 | 1 | |a Wisdom, Elias |4 aut | |
| 700 | 1 | |a Almoril-Porras, Agustin |4 aut | |
| 700 | 1 | |a Bowman, Ryan |4 aut | |
| 700 | 1 | |a Rodriguez, Tatiana |4 aut | |
| 700 | 1 | |a Adamson, Elise |4 aut | |
| 700 | 1 | |a Walder-Christensen, Kathryn K. |4 aut | |
| 700 | 1 | |a Hughes, Dalton |4 aut | |
| 700 | 1 | |a Schwennesen, Hannah |4 aut | |
| 700 | 1 | |a Mague, Stephen D. |4 aut | |
| 700 | 1 | |a Colón-Ramos, Daniel |4 aut | |
| 700 | 1 | |a Hultman, Rainbo |4 aut | |
| 700 | 1 | |a Bursac, Nenad |4 aut | |
| 700 | 1 | |a Dzirasa, Kafui |0 (orcid)0000-0003-4280-9994 |4 aut | |
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