Journal Article

Robotic microinjection enables large-scale transgenic studies of Caenorhabditis elegans

Bhatnagar, Sanjana, Li, Ang, Zuo, Runze, Saltzman, Arneet L., Li, Chengyin, Zhang, Pengsong, Liu, Xinyu, Qin, Zhen, Yu, Bin, Hung, Wesley, Pan, Peng, Pe, Karinna, Li, Ryan, Derry, W. Brent, Pilaka-Akella, Pallavi P., Sun, William, MacGillivray, Kailynn, Tang, Hua, Zhen, Mei, Xu, Pengfei, Zoberman, Michael, Wang, Shaojia, Martin, Charlotte, Calarco, John A., Premachandran, Sharanja, Zhang, Zefang

2024

Overview

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 . Manual injection, which remains low-throughput and labor-intensive, is a technical bottleneck for large-scale genetic studies of C. elegans. Here, the authors report a robotic microinjection system which facilitates injection speed while maintaining injection quality which is comparable to experienced experts.

• Publisher: Nature Publishing Group UK,Nature Publishing Group,Nature Portfolio
• Subject: 49/62 / 631/136/334/1582/712 / 631/1647/1511 / 631/1647/350/877 / 639/166/988 / 64/11 / Alternative Splicing / Animals / Animals, Genetically Modified / Caenorhabditis elegans / Caenorhabditis elegans - genetics / Caenorhabditis elegans Proteins - genetics / Caenorhabditis elegans Proteins - metabolism / Genetic screening / Genetically engineered organisms / Humanities and Social Sciences / Immobilization / Injection / Labor / Microfluidic devices / Microfluidics / Microinjection / Microinjections - instrumentation / Microinjections - methods / mRNA / multidisciplinary / Nematodes / Proteins / RNA, Messenger - genetics / RNA, Messenger - metabolism / RNA-binding protein / RNA-Binding Proteins - genetics / RNA-Binding Proteins - metabolism / Robotics / Robotics - instrumentation / Robotics - methods / Science / Science (multidisciplinary)