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Multiplex genome-editing in Arabidopsis thaliana using Mb3Cas12a

ABSTRACT: Multiplex genome-editing in Arabidopsis thaliana using Mb3Cas12a

PROVIDER: PRJNA683624 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR13220000_1.fastq.gz	Fastqsanger.gz
	SRR13220000_2.fastq.gz	Fastqsanger.gz
	SRR13220001_1.fastq.gz	Fastqsanger.gz
	SRR13220001_2.fastq.gz	Fastqsanger.gz
	SRR13220002_1.fastq.gz	Fastqsanger.gz

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Multiplex genome-editing in Arabidopsis thaliana using Mb3Cas12a

Project description:The use of CRISPR/Cas proteins for the creation of multiplex genome-engineering represents an important avenue for crop improvement, and further improvements for creation of knock-in plant lines via CRISPR-based technologies may enable the high-throughput creation of designer alleles. To circumvent limitations of the commonly used CRISPR/Cas9 system for multiplex genome-engineering, we explored the use of Moraxella bovoculi 3 Cas12a (Mb3Cas12a) for multiplex genome-editing in Arabidopsis thaliana. We identified optimized promoter sequences for driving expression of single transcript multiplex crRNA arrays in A. thaliana, resulting in stable germline transmission of Mb3Cas12a-edited alleles at multiple target sites. By utilizing this system, we demonstrate single-transcript multiplexed genome-engineering using of up to 13 crRNA targets. We further show high target specificity of Mb3Cas12a-based genome-editing via whole-genome sequencing. Taken together, our method provides a simplified platform for efficient multiplex-genome-engineering in plant-based systems.

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