Targeted Mutagenesis in Zea mays Using TALENs and the CRISPR/Cas System

Zhen Liang; Kang Zhang; Kunling Chen; Caixia Gao

doi:10.1016/j.jgg.2013.12.001

Volume 41 Issue 2

Feb. 2014

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Targeted Mutagenesis in Zea mays Using TALENs and the CRISPR/Cas System

doi: 10.1016/j.jgg.2013.12.001

State Key Laboratory of Plant Cell and Chromosome Engineering, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing 100101, China

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Corresponding author: E-mail address: cxgao@genetics.ac.cn (Caixia Gao)
Received Date: 2013-11-12
Accepted Date: 2013-12-05
Rev Recd Date: 2013-12-05

Available Online: 2013-12-14

Publish Date: 2014-02-20

Abstract

Abstract

Transcription activator-like effector nucleases (TALENs) and clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated (Cas) systems have emerged as powerful tools for genome editing in a variety of species. Here, we report, for the first time, targeted mutagenesis in Zea mays using TALENs and the CRISPR/Cas system. We designed five TALENs targeting 4 genes, namely ZmPDS, ZmIPK1A, ZmIPK, ZmMRP4, and obtained targeting efficiencies of up to 23.1% in protoplasts, and about 13.3% to 39.1% of the transgenic plants were somatic mutations. Also, we constructed two gRNAs targeting the ZmIPK gene in maize protoplasts, at frequencies of 16.4% and 19.1%, respectively. In addition, the CRISPR/Cas system induced targeted mutations in Z. mays protoplasts with efficiencies (13.1%) similar to those obtained with TALENs (9.1%). Our results show that both TALENs and the CRISPR/Cas system can be used for genome modification in maize.
- TAL-effector nucleases,
- CRISPR/Cas system,
- Knock-out

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References(28)

References

[1]	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
[2]	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
[3]	Chen, K., Gao, C. TALENs: customizable molecular DNA scissors for genome engineering of plants J. Genet. Genomics, 40 (2013),pp. 271-279
[4]	Cong, L., Ran, F.A., Cox, D. et al. Multiplex genome engineering using CRISPR/Cas systems Science, 339 (2013),pp. 819-823
[5]	Curtin, S.J., Zhang, F., Sander, J.D. et al. Targeted mutagenesis of duplicated genes in soybean with zinc-finger nucleases Plant Physiol., 156 (2011),pp. 466-473
[6]	Doyle, E.L., Booher, N.J., Standage, D.S. et al. TAL Effector-Nucleotide Targeter (TALE-NT) 2.0: tools for TAL effector design and target prediction Nucleic Acids Res., 40 (2012),pp. W117-W122
[7]	Fu, Y., Foden, J.A., Khayter, C. et al. High-frequency off-target mutagenesis induced by CRISPR-Cas nucleases in human cells Nat. Biotechnol., 31 (2013),pp. 822-826
[8]	Gaj, T., Gersbach, C.A., ZFN, TALEN, and CRISPR/Cas-based methods for genome engineering Trends Biotechnol., 31 (2013),pp. 397-405
[9]	Gasiunas, G., Barrangou, R., Horvath, P. et al. Cas9-crRNA ribonucleoprotein complex mediates specific DNA cleavage for adaptive immunity in bacteria Proc. Natl. Acad. Sci. USA, 109 (2012),pp. E2579-E2586
[10]	Ishida, Y., Hiei, Y., Komari, T. Agrobacterium-mediated transformation of maize Nat. Protocols, 2 (2007),pp. 1614-1621
[11]	Lei, Y., Guo, X., Deng, Y. et al. Cell Biosci., 3 (2013),p. 21
[12]	Li, J.F., Norville, J., Aach, J. et al. Nat. Biotechnol., 31 (2013),pp. 688-691
[13]	Li, T., Liu, B., Spalding, M.H. et al. High-efficiency TALEN-based gene editing produces disease-resistant rice Nat. Biotechnol., 30 (2012),pp. 390-392
[14]	Mali, P., Yang, L.H., Esvelt, K.M. et al. Science, 339 (2013),pp. 823-826
[15]	Nekrasov, V., S, B., Weigel, D. et al. Nat. Biotechnol., 31 (2013),pp. 691-693
[16]	Pattanayak, V., Lin, S., Guilinger, J.P. et al. High-throughput profiling of off-target DNA cleavage reveals RNA-programmed Cas9 nuclease specificity Nat. Biotechnol., 31 (2013),pp. 839-843
[17]	Qi, Y., Li, X., Zhang, Y. et al. G3 (Bethesda), 3 (2013),pp. 1707-1715
[18]	Shan, Q., Wang, Y., Chen, K. et al. Mol. Plant, 6 (2013),pp. 1365-1368
[19]	Shan, Q., Wang, Y., Li, J. et al. Targeted genome modification of crop plants using a CRISPR-Cas system Nat. Biotechnol., 31 (2013),pp. 686-688
[20]	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 (2012),pp. 76-81
[21]	Shi, J., Wang, H., Schellin, K. et al. Embryo-specific silencing of a transporter reduces phytic acid content of maize and soybean seeds Nat. Biotechnol., 25 (2007),pp. 930-937
[22]	Shi, J., Wang, H., Wu, Y. et al. Plant Physiol., 131 (2003),pp. 507-515
[23]	Shukla, V.K., Doyon, Y., Miller, J.C. et al. Nature, 459 (2009),pp. 437-441
[24]	Sun, Y., Thompson, M., Lin, G. et al. Inositol 1,3,4,5,6-pentakisphosphate 2-kinase from maize: molecular and biochemical characterization Plant Physiol., 144 (2007),pp. 1278-1291
[25]	Wang, Z., Li, J., Huang, H. et al. An integrated chip for the high-throughput synthesis of transcription activator-like effectors Angew. Chem. Int. Ed. Engl., 51 (2012),pp. 8505-8508
[26]	Wendt, T., Holm, P.B., Starker, C.G. et al. TAL effector nucleases induce mutations at a pre-selected location in the genome of primary barley transformants Plant Mol. Biol., 83 (2013),pp. 279-285
[27]	Zhang, F., Maeder, M.L., Unger-Wallace, E. et al. Proc. Natl. Acad. Sci. USA, 107 (2010),pp. 12028-12033
[28]	Zhang, Y., Zhang, F., Li, X. et al. Transcription activator-like effector nucleases enable efficient plant genome engineering Plant Physiol., 161 (2013),pp. 20-27