Generation of Inheritable A-to-G Transitions Using Adenine Base Editing and NG-PAM Cas9 in Arabidopsis thaliana

Up to 81% somatic A-to-G editing was achieved in primary transformants at a splice acceptor site with NGG PAM.

• Base editors were created by combining a modified Cas9 with adenosine deaminases to facilitate specific genetic changes.
• The highest efficiency of adenine base editing was observed with the vector pECNUS4, which carried the TadA8e enzyme.
• Simultaneous base editing of four homologs of NLR proteins was accomplished using a single guide RNA targeting a critical region.
• A-to-G edits were successfully fixed in three NLR genes across all targeted adenine sites, allowing for gene segregation in the next generation.
• The approach rescued an autoimmune phenotype in two generations by targeting multiple genes simultaneously.
• Editing on seven NLR genes was largely successful, with over 77% editing efficiency in six targets.

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