Expanding the scope of plant genome engineering with Cas12a orthologs and highly multiplexable editing systems

Yingxiao Zhang, Qiurong Ren, Xu Tang, Shishi Liu, Aimee A. Malzahn, Jianping Zhou, Jiaheng Wang, Desuo Yin, Changtian Pan, Mingzhu Yuan, Lan Huang, Han Yang, Yuxin Zhao, Qing Fang, Xuelian Zheng, Li Tian, Yanhao Cheng, Ysa Le, Bailey McCoy, Lidiya Franklin, Jeremy D. Selengut, Stephen M. Mount, Qiudeng Que, Yong Zhang () and Yiping Qi ()
Additional contact information
Yingxiao Zhang: University of Maryland
Qiurong Ren: University of Electronic Science and Technology of China
Xu Tang: University of Electronic Science and Technology of China
Shishi Liu: University of Electronic Science and Technology of China
Aimee A. Malzahn: University of Maryland
Jianping Zhou: University of Electronic Science and Technology of China
Jiaheng Wang: University of Electronic Science and Technology of China
Desuo Yin: University of Maryland
Changtian Pan: University of Maryland
Mingzhu Yuan: University of Electronic Science and Technology of China
Lan Huang: University of Electronic Science and Technology of China
Han Yang: University of Electronic Science and Technology of China
Yuxin Zhao: University of Electronic Science and Technology of China
Qing Fang: University of Electronic Science and Technology of China
Xuelian Zheng: University of Electronic Science and Technology of China
Li Tian: University of Electronic Science and Technology of China
Yanhao Cheng: University of Maryland
Ysa Le: University of Maryland
Bailey McCoy: University of Maryland
Lidiya Franklin: University of Maryland
Jeremy D. Selengut: University of Maryland
Stephen M. Mount: University of Maryland
Qiudeng Que: Syngenta
Yong Zhang: University of Electronic Science and Technology of China
Yiping Qi: University of Maryland

Nature Communications, 2021, vol. 12, issue 1, 1-11

Abstract: Abstract CRISPR-Cas12a is a promising genome editing system for targeting AT-rich genomic regions. Comprehensive genome engineering requires simultaneous targeting of multiple genes at defined locations. Here, to expand the targeting scope of Cas12a, we screen nine Cas12a orthologs that have not been demonstrated in plants, and identify six, ErCas12a, Lb5Cas12a, BsCas12a, Mb2Cas12a, TsCas12a and MbCas12a, that possess high editing activity in rice. Among them, Mb2Cas12a stands out with high editing efficiency and tolerance to low temperature. An engineered Mb2Cas12a-RVRR variant enables editing with more relaxed PAM requirements in rice, yielding two times higher genome coverage than the wild type SpCas9. To enable large-scale genome engineering, we compare 12 multiplexed Cas12a systems and identify a potent system that exhibits nearly 100% biallelic editing efficiency with the ability to target as many as 16 sites in rice. This is the highest level of multiplex edits in plants to date using Cas12a. Two compact single transcript unit CRISPR-Cas12a interference systems are also developed for multi-gene repression in rice and Arabidopsis. This study greatly expands the targeting scope of Cas12a for crop genome engineering.

Date: 2021
References: Add references at CitEc
Citations: View citations in EconPapers (1)

Downloads: (external link)
https://www.nature.com/articles/s41467-021-22330-w Abstract (text/html)

Related works:
This item may be available elsewhere in EconPapers: Search for items with the same title.

Export reference: BibTeX RIS (EndNote, ProCite, RefMan) HTML/Text

Persistent link: https://EconPapers.repec.org/RePEc:nat:natcom:v:12:y:2021:i:1:d:10.1038_s41467-021-22330-w

Ordering information: This journal article can be ordered from
https://www.nature.com/ncomms/

DOI: 10.1038/s41467-021-22330-w

Access Statistics for this article

Nature Communications is currently edited by Nathalie Le Bot, Enda Bergin and Fiona Gillespie

More articles in Nature Communications from Nature
Bibliographic data for series maintained by Sonal Shukla () and Springer Nature Abstracting and Indexing ().