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Oryza sativa Japonica Group

ABSTRACT: Precise integration of large DNA sequences in plant genomes using PrimeRoot editors

PROVIDER: PRJNA879048 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR23999171_1.fastq.gz	Fastqsanger.gz
	SRR23999171_2.fastq.gz	Fastqsanger.gz
	SRR23999172_1.fastq.gz	Fastqsanger.gz
	SRR23999172_2.fastq.gz	Fastqsanger.gz
	SRR23999173_1.fastq.gz	Fastqsanger.gz

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Project description:Adenine base editors (ABEs) with wide CRISPR compatibility and high activity improves the editing efficiency and arouses the off-target challenges as well. Here, we carried out a comprehensive evaluation of ABE8e and ABE9 induced DNA and RNA mutations in model organism rice. The whole-genome sequencing analysis on plants with rBE46b (SpCas9n-TadA8e), rBE49b (SpCas9n-TadA9), rBE50 (SpCas9n-NG-TadA8e), rBE53 (SpCas9n-NG-TadA9) reveals that the ABEs with TadA9 lead to a higher number of off-target A>G SNVs and ABEs with SpCas9n-NG lead to a higher total number of off-target SNVs. The analysis of T-DNAs (ABEs carrier) disclosed that the on-target mutations could happen before T-DNA integration to plant genomes as well as after T-DNA integration to plant genomes, while ABEs integrated into plant genomes lead to more A>G SNVs. We also detected off-target A>G RNA mutations in plants with higher expression of ABEs but not in plants with lower expression of ABEs. The off-target A>G RNA mutations tend to cluster while off-target A>G DNA mutations cluster in a very rare manner. The findings that CRISPRs, TadA variants, T-DNA integration, and ABE expression contribute ABEs’ specificity provide alternative ways to increase the specificity of ABEs.

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