Transgene-Free CRISPR-Edited Plants Indentification Using DsRED Fluorescence
November 13, 2019| |
Efficient removal of the editing machinery continues to be a concern in genome editing. To address this challenge, researchers at Universidad Politécnica de Valencia in Spain and Durham University in the UK used the modular cloning system Golden Braid and included a fluorescence-dependent transgene monitoring module to the genome-editing toolbox.
The technique was tested in tomato, rice, and Arabidopsis which showed that DsRED fluorescence visualization worked well in dry seeds as marker for the detection of the transgene. This shows that the method is an efficient way to select transgene-free dry seeds. In the first generation of Ds-RED-free CRISPR-Cas9 null segregants, gene editing of selected targets such as homozygous mutants was detected.
The results of the study indicate that the technique allows fast selection of transgene-free homozygous edited crop plants in a single generation after in vitro transformation.
Read the research article in Frontiers in Plant Science.
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