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Keywords

Bassoon, auxin, TIR1, Hair cells (HC)

Abstract

The Auxin Inducible Degron (AID) system is a conditional and reversible protein degradation system naturally occurring in plants. In the presence of the hormone Auxin, the AID tagged proteins are recruited into the plant’s E3 ubiquitin ligase machinery, ubiquitinated, and marked for degradation1. To adapt the AID system to the zebrafish model organism, our lab generated lines co-expressing Arabidopsis Thaliana plant TIR1 protein and GFP tagged with AID (GFP-AID) in superficially-located mechanosensory hair cells (HC) of the lateral line system and in spinal cord neurons found deeper in the body. Upon auxin application, GFP-AID expressed in the superficial HCs was degraded, while GFP-AID in the deeper spinal cord neurons was reduced by 50%. To increase degradation of AID tagged proteins in all tissues, we are currently working on improving the targeting vectors that use viral peptides (v2a) co-expressing TIR1 and a fluorescent protein. Furthermore, to show that this system can be used to degrade endogenous proteins, we are knocking in an AID tag in the genetic locus of the endogenous zebrafish Bassoon A gene. The synaptic cytomatrix protein, Bassoon, has been found in conventional synapses and ribbon synapses (present in mechanosensory hair cells of the auditory and vestibular system, in retinal photoreceptor and bipolar cells and in pinealocytes). In conventional synapses, Bassoon is shown to play an important role in the autophagy of synaptic proteins; however, this role has not been tested in ribbon synapses. By adapting the AID system to conditionally degrade Bassoon A in mechanosensory hair cells, we hope to 1) study its function in ribbon synapses, and 2) demonstrate the future potential of the AID system in the study of synaptic protein function in the zebrafish model organism.

Document Type

Poster

Publication Date

2024-03-21

Language

English

College

Life Sciences

Department

Neuroscience

University Standing at Time of Publication

Senior

Adapting the Auxin-Inducible Degron (AID) System to the Zebrafish Model Organism

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