Use of Fluorescent Protein Reporters for Assessing and Detecting Genome Editing Reagents and Transgene Expression in Plants

Research output: Chapter in Book/Report/Conference proceedingChapterpeer-review

1 Scopus citations

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., genotyping 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.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages115-127
Number of pages13
DOIs
StatePublished - 2023

Publication series

NameMethods in Molecular Biology
Volume2653
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Funding

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) Bioe-nergy 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.

FundersFunder number
Secure Ecosystem Engineering and Design
U.S. Department of Energy
Biological and Environmental Research
Oak Ridge National LaboratoryDE-AC05-00OR22725
Center for Bioenergy Innovation

    Keywords

    • Fluorescent protein
    • GFP
    • Genome editing
    • Plant transformation
    • eYGFPuv

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