Single-Base Editing in the Arabidopsis SUMO Conjugating Enzyme by Adenine Base Edition and Screening for a Rare Editing Event
摘要
SUMO conjugation is an essential process that regulates development and supports plant adaptation to environmental stress. We previously found that a Lys to Arg change in position 28 of the single-copy Arabidopsis thaliana SUMO conjugating enzyme SCE1 has increased in vitro activity, so we wanted to test the effect of this mutation in planta. The change is within reach for A to G base editing (AAG to AGG codon change) by an adenine deaminase-Cas9 fusion protein. We describe how we generated and identified two independent mutants carrying this change. The procedure may be useful for other cases of directed genome changes that occur at low frequency.