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

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

More Information

Corresponding author: E-mail address: byang@iastate.edu (Bing Yang)

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

摘要

- /
- /
- /
- /
- /
- /
- /
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.
注释:

HTML全文

参考文献(31)

[1]	Boch, J., Bonas, U. Ann. Rev. Phytopathol., 48 (2010),pp. 419-436
[2]	Boch, J., Scholze, H., Schornack, S. et al. Breaking the code of DNA binding specificity of TAL-type III effectors Science, 326 (2009),pp. 1509-1512
[3]	Cermak, T., Doyle, E.L., Christian, M. et al. Efficient design and assembly of custom TALEN and other TAL effector-based constructs for DNA targeting Nucleic Acids Res., 39 (2011),p. e82
[4]	Chipman, D., Barak, Z., Schloss, J.V. Biosynthesis of 2-aceto-2-hydroxy acids: acetolactate synthases and acetohydroxyacid synthases Biochim. Biophys. Acta, 1385 (1998),pp. 401-419
[5]	Christian, M., Cermak, T., Doyle, E.L. et al. Targeting DNA double-strand breaks with TAL effector nucleases Genetics, 186 (2010),pp. 757-761
[6]	Daxinger, L., Hunter, B., Sheikh, M. et al. Trends Plant Sci., 13 (2008),pp. 4-6
[7]	de Pater, S., Neuteboom, L.W., Pinas, J.E. et al. Plant Biotechnol. J., 7 (2009),pp. 821-835
[8]	de Pater, S., Pinas, J.E., Hooykaas, P.J. et al. Plant Biotechnol. J., 11 (2013),pp. 510-515
[9]	Endo, M., Osakabe, K., Ono, K. et al. Molecular breeding of a novel herbicide-tolerant rice by gene targeting Plant J., 52 (2007),pp. 157-166
[10]	Endo, M., Shimizu, T., Toki, S. Selection of transgenic rice plants using a herbicide tolerant form of the acetolactate synthase gene Methods Mol. Biol., 847 (2012),pp. 59-66
[11]	Endo, M., Mikami, M., Toki, S. Biallelic gene targeting in rice Plant Physiol., 170 (2016),pp. 667-677
[12]	Jinek, M., Chylinski, K., Fonfara, I. et al. A programmable dual-RNA-guided DNA endonuclease in adaptive bacterial immunity Science, 337 (2012),pp. 816-821
[13]	Kawai, K., Kaku, K., Izawa, N. et al. A novel mutant acetolactate synthase gene from rice cells, which confers resistance to ALS-inhibiting herbicides J. Pesticide Sci., 32 (2007),pp. 89-98
[14]	Li, J.F., Norville, J.E., Aach, J. et al. Nat. Biotechnol., 31 (2013),pp. 688-691
[15]	Li, T., Huang, S., Jiang, W.Z. et al. TAL nucleases (TALNs): hybrid proteins composed of TAL effectors and FokI DNA-cleavage domain Nucleic Acids Res., 39 (2011),pp. 359-372
[16]	Li, T., Liu, B., Chen, C.Y. et al. TALEN utilization in rice genome modifications Methods, 69 (2014),pp. 9-16
[17]	Mladenov, E., Iliakis, G. Induction and repair of DNA double strand breaks: the increasing spectrum of non-homologous end joining pathways Mutation Res., 711 (2011),pp. 61-72
[18]	Moscou, M.J., Bogdanove, A.J. A simple cipher governs DNA recognition by TAL effectors Science, 326 (2009),p. 1501
[19]	Murray, M.G., Thompson, W.F. Rapid isolation of high molecular weight plant DNA Nucleic Acids Res., 8 (1980),pp. 4321-4325
[20]	Osakabe, K., Endo, M., Kawai, K. et al. The mutant form of acetolactate synthase genomic DNA from rice is an efficient selectable marker for genetic transformation Mol. Breeding, 16 (2005),pp. 313-320
[21]	Reyon, D., Tsai, S.Q., Khayter, C. et al. FLASH assembly of TALENs for high-throughput genome editing Nat. Biotechnol., 30 (2012),pp. 460-465
[22]	San Filippo, J., Sung, P., Klein, H. Mechanism of eukaryotic homologous recombination Ann. Rev. Biochem., 77 (2008),pp. 229-257
[23]	Schmid-Burgk, J.L., Schmidt, T., Kaiser, V. et al. A ligation-independent cloning technique for high-throughput assembly of transcription activator-like effector genes Nat. Biotechnol., 31 (2013),pp. 76-81
[24]	Shimizu, T., Kaku, K., Kawai, K. et al.
[25]	Smith, J., Bibikova, M., Whitby, F.G. et al. Requirements for double-strand cleavage by chimeric restriction enzymes with zinc finger DNA-recognition domains Nucleic Acids Res., 28 (2000),pp. 3361-3369
[26]	Sun, Y., Zhang, X., Wu, C. et al. Engineering herbicide resistant rice plants through CRISPR/Cas9-mediated homologous recombination of the acetolactate synthase Mol. Plant, 9 (2016),pp. 628-631
[27]	Svitashev, S., Young, J.K., Schwartz, C. et al. Targeted mutagenesis, precise gene editing, and site-specific gene insertion in maize using Cas9 and guide RNA Plant Physiol., 169 (2015),pp. 931-945
[28]	Townsend, J.A., Wright, D.A., Winfrey, R.J. et al. High-frequency modification of plant genes using engineered zinc-finger nucleases Nature, 459 (2009),pp. 442-445
[29]	Weeks, D.P., Spalding, M.H., Yang, B. Use of designer nucleases for targeted gene and genome editing in plants Plant Biotechnol. J., 14 (2016),pp. 483-495
[30]	White, F.F., Potnis, N., Jones, J.B. et al. Mol. Plant Pathol., 10 (2009),pp. 749-766
[31]	Wright, D.A., Townsend, J.A., , Irwin, P.A. et al. High-frequency homologous recombination in plants mediated by zinc-finger nucleases Plant J., 44 (2005),pp. 693-705