Reference - Detail
| RRC ID | 84096 |
|---|---|
| Author | Pan P, Zoberman M, Zhang P, Premachandran S, Bhatnagar S, Pilaka-Akella PP, Sun W, Li C, Martin C, Xu P, Zhang Z, Li R, Hung W, Tang H, MacGillivray K, Yu B, Zuo R, Pe K, Qin Z, Wang S, Li A, Derry WB, Zhen M, Saltzman AL, Calarco JA, Liu X. |
| Title | Robotic microinjection enables large-scale transgenic studies of Caenorhabditis elegans. |
| Journal | Nat Commun |
| Abstract |
The nematode Caenorhabditis elegans is widely employed as a model organism to study basic biological mechanisms. However, transgenic C. elegans are generated by manual injection, which remains low-throughput and labor-intensive, limiting the scope of approaches benefitting from large-scale transgenesis. Here, we report a robotic microinjection system, integrating a microfluidic device capable of reliable worm immobilization, transfer, and rotation, for high-speed injection of C. elegans. The robotic system provides an injection speed 2-3 times faster than that of experts with 7-22 years of experience while maintaining comparable injection quality and only limited trials needed by users to become proficient. We further employ our system in a large-scale reverse genetic screen using multiplexed alternative splicing reporters, and find that the TDP-1 RNA-binding protein regulates alternative splicing of zoo-1 mRNA, which encodes variants of the zonula occludens tight junction proteins. With its high speed, high accuracy, and high efficiency in worm injection, this robotic system shows great potential for high-throughput transgenic studies of C. elegans. |
| Volume | 15(1) |
| Pages | 8848 |
| Published | 2024-10-14 |
| DOI | 10.1038/s41467-024-53108-5 |
| PII | 10.1038/s41467-024-53108-5 |
| PMID | 39397017 |
| PMC | PMC11471809 |
| MeSH | Alternative Splicing Animals Animals, Genetically Modified* Caenorhabditis elegans* / genetics Caenorhabditis elegans Proteins* / genetics Caenorhabditis elegans Proteins* / metabolism Microinjections* / instrumentation Microinjections* / methods RNA, Messenger / genetics RNA, Messenger / metabolism RNA-Binding Proteins / genetics RNA-Binding Proteins / metabolism Robotics* / instrumentation Robotics* / methods |
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| The most frequently cited source | X(Twitter) |
| Total number of mentions | 12 |
| Altmetric score changes over past 6months | 0.0 |
| Resource | |
| C.elegans | tm668 |