A simple method to dramatically increase C. elegans germline microinjection efficiency.

Autor: Gibney TV; Department of Biology, University of Virginia, Charlottesville, VA, USA. Electronic address: tg7uw@virginia.edu., Favichia M; Department of Biology, University of Virginia, Charlottesville, VA, USA., Latifi L; Department of Biology, University of Virginia, Charlottesville, VA, USA., Medwig-Kinney TN; Department of Biochemistry & Cell Biology, Stony Brook University, Stony Brook, NY, USA., Matus DQ; Department of Biochemistry & Cell Biology, Stony Brook University, Stony Brook, NY, USA., McIntyre DC; Department of Biology, University of Virginia, Charlottesville, VA, USA., Arrigo AB; Department of Biology, University of Virginia, Charlottesville, VA, USA., Branham KR; Department of Biology, University of Virginia, Charlottesville, VA, USA., Bubrig LT; Department of Biology, University of Virginia, Charlottesville, VA, USA., Ghaddar A; Department of Biology, University of Virginia, Charlottesville, VA, USA., Jiranek JA; Department of Biology, University of Virginia, Charlottesville, VA, USA., Liu KE; Department of Cell Biology, University of Virginia, Charlottesville, VA, USA., Marcucci CG; Neuroscience Graduate Program, University of Virginia, Charlottesville, VA, USA., Porter RJ; Department of Biology, University of Virginia, Charlottesville, VA, USA., Pani AM; Department of Biology, University of Virginia, Charlottesville, VA, USA; Department of Cell Biology, University of Virginia, Charlottesville, VA, USA. Electronic address: amp2na@virginia.edu.
Jazyk: angličtina
Zdroj: Developmental biology [Dev Biol] 2023 Oct; Vol. 502, pp. 63-67. Date of Electronic Publication: 2023 Jul 09.
DOI: 10.1016/j.ydbio.2023.07.003
Abstrakt: 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.
(Copyright © 2023 The Authors. Published by Elsevier Inc. All rights reserved.)
Databáze: MEDLINE
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