期刊
PLANT BIOTECHNOLOGY JOURNAL
卷 -, 期 -, 页码 -出版社
WILEY
DOI: 10.1111/pbi.14122
关键词
deletion; genome editing; CRISPR; class I; type I-C CRISPR system
We developed a new Dvu I-C system for plant genome editing, which can efficiently achieve controllable large fragment deletions with an editing efficiency of up to 86.67%. This system is sensitive to spacer length and mismatch, benefiting target specificity.
The CRISPR-Cas systems have been widely used as genome editing tools, with type II and V systems typically introducing small indels, and type I system mediating long-range deletions. However, the precision of type I systems for large fragment deletion is still remained to be optimized. Here, we developed a compact Cascade-Cas3 Dvu I-C system with Cas11c for plant genome editing. The Dvu I-C system was efficient to introduce controllable large fragment deletion up to at least 20 kb using paired crRNAs. The paired-crRNAs design also improved the controllability of deletions for the type I-E system. Dvu I-C system was sensitive to spacer length and mismatch, which was benefit for target specificity. In addition, we showed that the Dvu I-C system was efficient for generating stable transgenic lines in maize and rice with the editing efficiency up to 86.67%. Overall, Dvu I-C system we developed here is powerful for achieving controllable large fragment deletions.
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