Dataset's files
| Action | DRS | |||
|---|---|---|---|---|
| SRR18513500_1.fastq.gz | Fastqsanger.gz | |||
| SRR18513500_2.fastq.gz | Fastqsanger.gz | |||
| SRR18513600_1.fastq.gz | Fastqsanger.gz | |||
| SRR18513600_2.fastq.gz | Fastqsanger.gz | |||
| SRR18513700_1.fastq.gz | Fastqsanger.gz |
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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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