Campus Units

Genetics, Development and Cell Biology

Document Type

Article

Publication Date

9-2014

Journal or Book Title

Nucleic Acids Research

Volume

42

Issue

17

First Page

10903

Last Page

10914

DOI

10.1093/nar/gku806

Abstract

The Cas9/sgRNA of the CRISPR/Cas system has emerged as a robust technology for targeted gene editing in various organisms, including plants, where Cas9/sgRNA-mediated small deletions/insertions at single cleavage sites have been reported in transient and stable transformations, although genetic transmission of edits has been reported only in Arabidopsisand rice. Large chromosomal excision between two remote nuclease-targeted loci has been reported only in a few non-plant species. Here we report in rice Cas9/sgRNA-induced large chromosomal segment deletions, the inheritance of genome edits in multiple generations and construction of a set of facile vectors for high-efficiency, multiplex gene targeting. Four sugar efflux transporter genes were modified in rice at high efficiency; the most efficient system yielding 87–100% editing in T0 transgenic plants, all with di-allelic edits. Furthermore, genetic crosses segregating Cas9/sgRNA transgenes away from edited genes yielded several genome-edited but transgene-free rice plants. We also demonstrated proof-of-efficiency of Cas9/sgRNAs in producing large chromosomal deletions (115–245 kb) involving three different clusters of genes in rice protoplasts and verification of deletions of two clusters in regenerated T0 generation plants. Together, these data demonstrate the power of our Cas9/sgRNA platform for targeted gene/genome editing in rice and other crops, enabling both basic research and agricultural applications.

Comments

This article is published as Zhou, Huanbin, Bo Liu, Donald P. Weeks, Martin H. Spalding, and Bing Yang. "Large chromosomal deletions and heritable small genetic changes induced by CRISPR/Cas9 in rice." Nucleic acids research 42, no. 17 (2014): 10903-10914. 10.1093/nar/gku806

Creative Commons License

Creative Commons Attribution-Noncommercial 4.0 License
This work is licensed under a Creative Commons Attribution-Noncommercial 4.0 License

Copyright Owner

The Authors

Language

en

File Format

application/pdf

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