A simple method to dramatically increase C. elegans germline microinjection efficiency
A simple method to dramatically increase C. elegans germline microinjection efficiency
Abstract Genome manipulation methods in C. elegans require microinjecting DNA or ribonucleoprotein complexes into the microscopic core of the gonadal syncytium. These microinjections are technically demanding and represent a key bottleneck for all genome engineering and transgenic approaches in C. elegans. While there have been steady improvements in the ease and efficiency of genetic methods for C. elegans genome manipulation, there have not been comparable advances in the physical process of microinjection. Here, we report a simple and inexpensive method for handling worms using a paintbrush during the injection process that nearly tripled average microinjection rates compared to traditional worm handling methods. We found that the paintbrush increased injection throughput by substantially increasing both injection speeds and post-injection survival rates. In addition to dramatically and universally increasing injection efficiency for experienced personnel, the paintbrush method also significantly improved the abilities of novice investigators to perform key steps in the microinjection process. We expect that this method will benefit the C. elegans community by increasing the speed at which new strains can be generated and will also make microinjection-based approaches less challenging and more accessible to personnel and labs without extensive experience.
Favichia Michelle、Arrigo Angelo B.、Ghaddar Abbas、Pani Ariel M.、Porter Robert J.、Medwig-Kinney Taylor N.、Bubrig Louis T.、Branham Kendall R.、Gibney Theresa V.、Jiranek Juliana A.、McIntyre Daniel C.、Matus David Q.、Marcucci Charles G.、Liu Kendra E.、Latifi Laila
Department of Biology, University of VirginiaDepartment of Biology, University of VirginiaDepartment of Biology, University of VirginiaDepartment of Biology, University of Virginia||Department of Cell Biology, University of VirginiaDepartment of Biology, University of VirginiaDepartment of Biochemistry & Cell Biology, Stony Brook UniversityDepartment of Biology, University of VirginiaDepartment of Biology, University of VirginiaDepartment of Biology, University of VirginiaDepartment of Biology, University of VirginiaDepartment of Biology, University of VirginiaDepartment of Biochemistry & Cell Biology, Stony Brook University||David Matus is a paid consultant of Arcadia ScienceNeuroscience Graduate Program, University of VirginiaNeuroscience Graduate Program, University of VirginiaDepartment of Biology, University of Virginia
遗传学生物科学研究方法、生物科学研究技术分子生物学
C. elegansmicroinjectiongenome engineeringgene editingCRISPRtransgenesis
Favichia Michelle,Arrigo Angelo B.,Ghaddar Abbas,Pani Ariel M.,Porter Robert J.,Medwig-Kinney Taylor N.,Bubrig Louis T.,Branham Kendall R.,Gibney Theresa V.,Jiranek Juliana A.,McIntyre Daniel C.,Matus David Q.,Marcucci Charles G.,Liu Kendra E.,Latifi Laila.A simple method to dramatically increase C. elegans germline microinjection efficiency[EB/OL].(2025-03-28)[2025-06-06].https://www.biorxiv.org/content/10.1101/2023.03.23.533855.点此复制
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