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Genome-wide specificity of prime editors in plants
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Genome-wide specificity of prime editors in plants
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Journal Article
Genome-wide specificity of prime editors in plants
2021
Overview
Although prime editors (PEs) have the potential to facilitate precise genome editing in therapeutic, agricultural and research applications, their specificity has not been comprehensively evaluated. To provide a systematic assessment in plants, we first examined the mismatch tolerance of PEs in plant cells and found that the editing frequency was influenced by the number and location of mismatches in the primer binding site and spacer of the prime editing guide RNA (pegRNA). Assessing the activity of 12 pegRNAs at 179 predicted off-target sites, we detected only low frequencies of off-target edits (0.00~0.23%). Whole-genome sequencing of 29 PE-treated rice plants confirmed that PEs do not induce genome-wide pegRNA-independent off-target single-nucleotide variants or small insertions/deletions. We also show that ectopic expression of the Moloney murine leukemia virus reverse transcriptase as part of the PE does not change retrotransposon copy number or telomere structure or cause insertion of pegRNA or messenger RNA sequences into the genome.
A very high specificity of prime editors is demonstrated in a study of their off-target activity in plants.
Publisher
Nature Publishing Group US,Nature Publishing Group
Subject
/ 631/449
/ 631/61
/ Biomedical and Life Sciences
/ Biomedical Engineering/Biotechnology
/ Editing
/ Genome-wide association studies
/ Genomes
/ Leukemia
/ Methods
/ Moloney murine leukemia virus - genetics
/ mRNA
/ Mutation
/ Reverse Transcription - genetics
