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 language | English |
|---|---|
| Title of host publication | Methods in Molecular Biology |
| Publisher | Humana Press Inc. |
| Pages | 115-127 |
| Number of pages | 13 |
| DOIs | |
| State | Published - 2023 |
Publication series
| Name | Methods in Molecular Biology |
|---|---|
| Volume | 2653 |
| 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.
Keywords
- Fluorescent protein
- GFP
- Genome editing
- Plant transformation
- eYGFPuv