Abstract
Fluorescent protein reporters have been widely used for monitoring the expression of target genes in various engineered organisms. Although a wide range of analytical approaches (e.g., genoty** PCR, digital PCR, DNA sequencing) have been utilized to detect and identify genome editing reagents and transgene expression in genetically modified plants, these methods are usually limited to use in the late stages of plant transformation and can only be used invasively. Here we describe GFP- and eYGFPuv-based strategies and methods for assessing and detecting genome editing reagents and transgene expression in plants, including protoplast transformation, leaf infiltration, and stable transformation. These methods and strategies enable easy, noninvasive screening of genome editing and transgenic events in plants.
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Acknowledgments
The authors are grateful to C.A. Eckert (Oak Ridge National Laboratory, Oak Ridge, TN, USA) for editing the manuscript. The writing of this manuscript was supported by the Center for Bioenergy Innovation, a US Department of Energy (DOE) Bioenergy Research Center supported by the Biological and Environmental Research (BER) program, and the US DOE BER Genomic Science Program, as part of the Secure Ecosystem Engineering and Design (SEED) Scientific Focus Area. Oak Ridge National Laboratory is managed by UT-Battelle, LLC for the US Department of Energy under Contract Number DE-AC05-00OR22725.
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This manuscript has been authored by UT-Battelle, LLC under Contract No. DE-AC05-00OR22725 with the US Department of Energy. The US Government retains and the publisher, by accepting the article for publication, acknowledges that the US Government retains a nonexclusive, paid-up, irrevocable, worldwide license to publish or reproduce the published form of this manuscript, or allow others to do so, for the US Government purposes. The Department of Energy will provide public access to these results of federally sponsored research in accordance with the DOE Public Access Plan (http://energy.gov/downloads/doe-public-access-plan).
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Yuan, G., Tuskan, G.A., Yang, X. (2023). Use of Fluorescent Protein Reporters for Assessing and Detecting Genome Editing Reagents and Transgene Expression in Plants. In: Yang, B., Harwood, W., Que, Q. (eds) Plant Genome Engineering. Methods in Molecular Biology, vol 2653. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3131-7_8
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DOI: https://doi.org/10.1007/978-1-0716-3131-7_8
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