Circumventing neural damage in a C. elegans chemosensory circuit using genetically engineered synapses

Rabinowitch, Ithai; Upadhyaya, Bishal; Pant, Aaradhya; Galski, Dolev; Kreines, Lena; Bai, Jihong

doi:10.1016/j.cels.2020.12.003

Cited by 3 publications

(6 citation statements)

References 53 publications

(75 reference statements)

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“…Moreover, several groups have established that selectively expressing Cx36 is sufficient to reconstitute a functional electrical synapse between two connected neurons. Specifically, the presence and function of this resultant Cx36/Cx36 synapse has been confirmed via microscopy (Rabinowitch et al ., 2014), measurements of synaptic physiology (Rabinowitch et al ., 2014), calcium imaging (Hawk et al ., 2018), and behavior (Choi et al ., 2020; Hawk et al ., 2018; Rabinowitch, 2022; Rabinowitch et al ., 2014; Rabinowitch and Schafer, 2015; Rabinowitch et al ., 2021). Thus, we chose to benchmark our new connexin pair in C. elegans against the well characterized Cx36/Cx36 synapse.…”

Section: Resultsmentioning

confidence: 99%

“…As such, ectopic expression of the vertebrate connexin36 (Cx36) across two connected C. elegans neurons results in the formation of an electrical synapse that is inert to endogenous C. elegans gap junction proteins (i.e., innexins). Previously work with this approach has successfully been used to modify circuit physiology and multiple behavioral contexts including migration of C. elegans in response to various chemical and temperature cues (Choi et al ., 2020; Hawk et al ., 2018; Rabinowitch et al ., 2014; Rabinowitch et al, 2021).…”

Section: Introductionmentioning

confidence: 99%

“…The transformative potential for using gap junctions to repair dysfunctional circuits has been advanced in C. elegans as well (Rabinowitch, 2022; Rabinowitch et al ., 2021). There, circuit editing was utilized to restore normal behavior in animals with circuit impairments.…”

Section: Introductionmentioning

confidence: 99%

“…Nevertheless, this work also highlighted a significant challenge to employing gap junctions to edit select circuits in higher-complexity organisms. Specifically, when Cx36 was expressed in two neurons of the same cell class in C. elegans , AWC-left and AWC-right, animals showed altered behavior in response to olfactory cues (Rabinowitch et al ., 2021). Thus, the feature of connexins to form single-isoform homotypic gap junctions dramatically limits its potential to selectively edit brain circuits in vertebrate brains.…”

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Long-term precision editing of neural circuits in mammals using engineered gap junction hemichannels

Ransey¹,

Chesnov²,

Wisdom

et al. 2021

Preprint

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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 circuit, while leaving the activity of the presynaptic brain cell undisturbed remain sparce. To address this gap, we developed a novel class of electrical synapses by selectively engineering two connexin proteins found in Morone americana (white perch fish): connexin34.7 (Cx34.7) and connexin35 (Cx35). By iteratively exploiting protein mutagenesis, a novel in vitro assay of connexin docking, and computational modeling of connexin hemichannel interactions, we uncovered the pattern of structural motifs that broadly determine connexin hemichannel docking. We then utilized this knowledge to design Cx34.7 and Cx35 hemichannels that dock with each other, but not with themselves nor other major connexins expressed in the human central nervous system. We validated these hemichannels in vivo, demonstrating that they facilitate communication between two neurons in Caenorhabditis elegans and recode a learned behavioral preference. This system can be applied to edit circuits composed by pairs of genetically defined brain cell types across multiple species. Thus, we establish a potentially translational approach, Long-term integration of Circuits using connexins (LinCx), for context-precise circuit-editing with unprecedented spatiotemporal specificity.

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Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Long-term precision editing of neural circuits in mammals using engineered gap junction hemichannels

Ransey¹,

Chesnov²,

Wisdom

et al. 2021

Preprint

View full text Add to dashboard Cite

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Synthetic biology in multicellular organisms: Opportunities in nematodes

Kukhtar

Fussenegger

2023

Biotech & Bioengineering

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Synthetic biology has mainly focused on introducing new or altered functionality in single cell systems: primarily bacteria, yeast, or mammalian cells. Here, we describe the extension of synthetic biology to nematodes, in particular the well‐studied model organism Caenorhabditis elegans, as a convenient platform for developing applications in a multicellular setting. We review transgenesis techniques for nematodes, as well as the application of synthetic biology principles to construct nematode gene switches and genetic devices to control motility. Finally, we discuss potential applications of engineered nematodes.

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