Robotic microinjection enables large-scale transgenic studies of Caenorhabditis elegans

Peng Pan, Michael Zoberman, Pengsong Zhang, Sharanja Premachandran, Sanjana Bhatnagar, Pallavi P. Pilaka-Akella, William Sun, Chengyin Li, Charlotte Martin, Pengfei Xu, Zefang Zhang, Ryan Li, Wesley Hung, Hua Tang, Kailynn MacGillivray, Bin Yu, Runze Zuo, Karinna Pe, Zhen Qin, Shaojia Wang, Ang Li, W. Brent Derry, Mei Zhen, Arneet L. Saltzman, John A. Calarco () and Xinyu Liu ()
Additional contact information
Peng Pan: University of Toronto
Michael Zoberman: University of Toronto
Pengsong Zhang: University of Toronto
Sharanja Premachandran: University of Toronto
Sanjana Bhatnagar: University of Toronto
Pallavi P. Pilaka-Akella: University of Toronto
William Sun: Upper Canada College
Chengyin Li: University of Toronto
Charlotte Martin: University of Toronto
Pengfei Xu: University of Toronto
Zefang Zhang: University of Toronto
Ryan Li: University of Toronto
Wesley Hung: 600 University Ave
Hua Tang: University of Toronto
Kailynn MacGillivray: University of Toronto
Bin Yu: The Hospital for Sick Children
Runze Zuo: University of Toronto
Karinna Pe: University of Toronto
Zhen Qin: University of Toronto
Shaojia Wang: University of Toronto
Ang Li: University of Toronto
W. Brent Derry: The Hospital for Sick Children
Mei Zhen: 600 University Ave
Arneet L. Saltzman: University of Toronto
John A. Calarco: University of Toronto
Xinyu Liu: University of Toronto

Nature Communications, 2024, vol. 15, issue 1, 1-15

Abstract: 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.

Date: 2024
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DOI: 10.1038/s41467-024-53108-5

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