Robotic microinjection enables large-scale transgenic studies of Caenorhabditis elegans.

Autor:	Pan P; Department of Mechanical and Industrial Engineering, University of Toronto, Toronto, ON, Canada.; Department of Mechanical Engineering, McGill University, Montreal, QC, Canada., Zoberman M; Department of Cell & Systems Biology, University of Toronto, Toronto, ON, Canada., Zhang P; Department of Mechanical and Industrial Engineering, University of Toronto, Toronto, ON, Canada., Premachandran S; Department of Cell & Systems Biology, University of Toronto, Toronto, ON, Canada., Bhatnagar S; Department of Cell & Systems Biology, University of Toronto, Toronto, ON, Canada., Pilaka-Akella PP; Department of Cell & Systems Biology, University of Toronto, Toronto, ON, Canada., Sun W; Upper Canada College, Toronto, ON, Canada., Li C; Department of Cell & Systems Biology, University of Toronto, Toronto, ON, Canada., Martin C; Department of Cell & Systems Biology, University of Toronto, Toronto, ON, Canada., Xu P; Department of Mechanical and Industrial Engineering, University of Toronto, Toronto, ON, Canada., Zhang Z; Department of Mechanical and Industrial Engineering, University of Toronto, Toronto, ON, Canada., Li R; Department of Mechanical and Industrial Engineering, University of Toronto, Toronto, ON, Canada., Hung W; Lunenfeld-Tanenbaum Research Institute, Mount Sinai Hospital, 600 University Ave, Toronto, ON, Canada., Tang H; Department of Cell & Systems Biology, University of Toronto, Toronto, ON, Canada., MacGillivray K; Department of Cell & Systems Biology, University of Toronto, Toronto, ON, Canada., Yu B; Developmental and Stem Cell Biology, The Hospital for Sick Children, Toronto, ON, Canada., Zuo R; Department of Mechanical and Industrial Engineering, University of Toronto, Toronto, ON, Canada., Pe K; Department of Cell & Systems Biology, University of Toronto, Toronto, ON, Canada., Qin Z; Department of Mechanical and Industrial Engineering, University of Toronto, Toronto, ON, Canada., Wang S; Department of Mechanical and Industrial Engineering, University of Toronto, Toronto, ON, Canada., Li A; Department of Mechanical and Industrial Engineering, University of Toronto, Toronto, ON, Canada., Derry WB; Developmental and Stem Cell Biology, The Hospital for Sick Children, Toronto, ON, Canada., Zhen M; Lunenfeld-Tanenbaum Research Institute, Mount Sinai Hospital, 600 University Ave, Toronto, ON, Canada., Saltzman AL; Department of Cell & Systems Biology, University of Toronto, Toronto, ON, Canada., Calarco JA; Department of Cell & Systems Biology, University of Toronto, Toronto, ON, Canada. john.calarco@utoronto.ca., Liu X; Department of Mechanical and Industrial Engineering, University of Toronto, Toronto, ON, Canada. xyliu@mie.utoronto.ca.; Institute of Biomedical Engineering, University of Toronto, Toronto, ON, Canada. xyliu@mie.utoronto.ca.
Jazyk:	angličtina
Zdroj:	Nature communications [Nat Commun] 2024 Oct 14; Vol. 15 (1), pp. 8848. Date of Electronic Publication: 2024 Oct 14.
DOI:	10.1038/s41467-024-53108-5
Abstrakt:	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. (© 2024. The Author(s).)
Databáze:	MEDLINE
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