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PMID: 24576457 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Targeted mutagenesis in Zea mays using TALENs and the CRISPR/Cas system.

Journal of genetics and genomics = Yi chuan xue bao ·Vol. 41 ·No. 2 ·2014-02-20 ·Pages 63-8

Liang Z, Zhang K, Chen K, Gao C

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.

Keywords
CRISPR/Cas system Knock-out TAL-effector nucleases Zea mays
MeSH Terms
Base Sequence CRISPR-Cas Systems/genetics Clustered Regularly Interspaced Short Palindromic Repeats/genetics DNA Restriction Enzymes/metabolism Genes, Plant/genetics Genetic Engineering/methods Genomics Mutagenesis, Site-Directed Mutation Plants, Genetically Modified Zea mays/genetics
Chemicals
DNA Restriction Enzymes
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Liang Zhen
State Key Laboratory of Plant Cell and Chromosome Engineering, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing 100101, China.
Zhang Kang
State Key Laboratory of Plant Cell and Chromosome Engineering, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing 100101, China.
Chen Kunling
State Key Laboratory of Plant Cell and Chromosome Engineering, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing 100101, China.
Gao Caixia
State Key Laboratory of Plant Cell and Chromosome Engineering, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing 100101, China. Electronic address: cxgao@genetics.ac.cn.
Article Info
Journal
Journal of genetics and genomics = Yi chuan xue bao
Abbr.
J Genet Genomics
ISSN
1673-8527
Published
2014-02-20
Epub
2013-00-14
Pages
63-8
Language
English
Region
China
NLM ID
101304616
Subset
IM
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