TALEN-Mediated Homologous Recombination Produces Site-Directed DNA Base Change and Herbicide-Resistant Rice

Ting Li; Bo Liu; Chih Ying Chen; Bing Yang

doi:10.1016/j.jgg.2016.03.005

Volume 43 Issue 5

May 2016

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Article Navigation > Journal of Genetics and Genomics > 2016 > 43(5): 297-305

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TALEN-Mediated Homologous Recombination Produces Site-Directed DNA Base Change and Herbicide-Resistant Rice

doi: 10.1016/j.jgg.2016.03.005

Department of Genetics, Development and Cell Biology, Iowa State University, Ames 50011, USA

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Corresponding author: E-mail address: byang@iastate.edu (Bing Yang)
Received Date: 2016-01-30
Accepted Date: 2016-03-09
Rev Recd Date: 2016-03-09

Available Online: 2016-03-22

Publish Date: 2016-05-20

Abstract

Abstract

Over the last decades, much endeavor has been made to advance genome editing technology due to its promising role in both basic and synthetic biology. The breakthrough has been made in recent years with the advent of sequence-specific endonucleases, especially zinc finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs) and clustered regularly interspaced short palindromic repeats (CRISPRs) guided nucleases (e.g., Cas9). In higher eukaryotic organisms, site-directed mutagenesis usually can be achieved through non-homologous end-joining (NHEJ) repair to the DNA double-strand breaks (DSBs) caused by the exogenously applied nucleases. However, site-specific gene replacement or genuine genome editing through homologous recombination (HR) repair to DSBs remains a challenge. As a proof of concept gene replacement through TALEN-based HR in rice (Oryza sativa), we successfully produced double point mutations in rice acetolactate synthase gene (OsALS) and generated herbicide resistant rice lines by using TALENs and donor DNA carrying the desired mutations. After ballistic delivery into rice calli of TALEN construct and donor DNA, nine HR events with different genotypes ofOsALS were obtained in T₀ generation at the efficiency of 1.4%–6.3% from three experiments. The HR-mediated gene edits were heritable to the progeny of T₁ generation. The edited T₁ plants were as morphologically normal as the control plants while displayed strong herbicide resistance. The results demonstrate the feasibility of TALEN-mediated genome editing in rice and provide useful information for further genome editing by other nuclease-based genome editing platforms.
- TALEN,
- Genome editing,
- Homologous recombination,
- Gene replacement,
- Site-directed mutagenesis,
- Acetolactate synthase,
- Herbicide resistance,
- Rice

Present address: Division of Biology and Biological Engineering, California Institute of Technology, Pasadena, USA.

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References(31)

References

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