Precise base editing in rice, wheat and maize with a Cas9-cytidine deaminase fusion

Targeted base editing in plants without the need for a foreign DNA donor or double-stranded DNA cleavage would accelerate genome modification and breeding in a wide array of crops. We used a CRISPR–Cas9 nickase-cytidine deaminase fusion to achieve targeted conversion of cytosine to thymine from position 3 to 9 within the protospacer in both protoplasts and regenerated rice, wheat and maize plants at frequencies of up to 43.48%.

[1]  C. Topp,et al.  Centromeric Retroelements and Satellites Interact with Maize Kinetochore Protein CENH3 , 2002, The Plant Cell Online.

[2]  S. Henikoff,et al.  Single-nucleotide mutations for plant functional genomics. , 2003, Annual review of plant biology.

[3]  S. Henikoff,et al.  TILLING. Traditional Mutagenesis Meets Functional Genomics , 2004, Plant Physiology.

[4]  S. Fuerstenberg,et al.  A reverse genetic, nontransgenic approach to wheat crop improvement by TILLING , 2005, Nature Biotechnology.

[5]  N. Wu,et al.  Molecular analysis of lipoxygenase (LOX) genes in common wheat and phylogenetic investigation of LOX proteins from model and crop plants , 2010 .

[6]  Mark H. Wright,et al.  Genome-wide association mapping reveals a rich genetic architecture of complex traits in Oryza sativa , 2011, Nature communications.

[7]  I. Szarejko,et al.  TILLING - a shortcut in functional genomics , 2011, Journal of Applied Genetics.

[8]  Jun Li,et al.  Targeted genome modification of crop plants using a CRISPR-Cas system , 2013, Nature Biotechnology.

[9]  Kang Zhang,et al.  Targeted mutagenesis in Zea mays using TALENs and the CRISPR/Cas system. , 2014, Journal of genetics and genomics = Yi chuan xue bao.

[10]  Jin-Soo Kim,et al.  Cas-OFFinder: a fast and versatile algorithm that searches for potential off-target sites of Cas9 RNA-guided endonucleases , 2014, Bioinform..

[11]  Yanpeng Wang,et al.  Simultaneous editing of three homoeoalleles in hexaploid bread wheat confers heritable resistance to powdery mildew , 2014, Nature Biotechnology.

[12]  Daniel F. Voytas,et al.  Precision Genome Engineering and Agriculture: Opportunities and Regulatory Challenges , 2014, PLoS biology.

[13]  E. Lander,et al.  Development and Applications of CRISPR-Cas9 for Genome Engineering , 2014, Cell.

[14]  Hui-Li Xing,et al.  A CRISPR/Cas9 toolkit for multiplex genome editing in plants , 2014, BMC Plant Biology.

[15]  D. Voytas,et al.  High-frequency, precise modification of the tomato genome , 2015, Genome Biology.

[16]  Joshua K Young,et al.  Targeted Mutagenesis, Precise Gene Editing, and Site-Specific Gene Insertion in Maize Using Cas9 and Guide RNA[OPEN] , 2015, Plant Physiology.

[17]  A. Kondo,et al.  Targeted nucleotide editing using hybrid prokaryotic and vertebrate adaptive immune systems , 2016, Science.

[18]  D. Guo,et al.  Single base substitution in OsCDC48 is responsible for premature senescence and death phenotype in rice , 2015, Journal of integrative plant biology.

[19]  David R. Liu,et al.  Programmable editing of a target base in genomic DNA without double-stranded DNA cleavage , 2016, Nature.

[20]  Gaelen T. Hess,et al.  Directed evolution using dCas9-targeted somatic hypermutation in mammalian cells , 2016, Nature Methods.

[21]  Yi Zhang,et al.  Efficient and transgene-free genome editing in wheat through transient expression of CRISPR/Cas9 DNA or RNA , 2016, Nature Communications.

[22]  Yan Song,et al.  Targeted AID-mediated mutagenesis (TAM) enables efficient genomic diversification in mammalian cells , 2016, Nature Methods.

[23]  Honghui Lin,et al.  A CRISPR/Cas9 toolkit for efficient targeted base editing to induce genetic variations in rice , 2017, Science China Life Sciences.

[24]  Jian‐Kang Zhu,et al.  Precise Editing of a Target Base in the Rice Genome Using a Modified CRISPR/Cas9 System. , 2017, Molecular plant.

[25]  Yunde Zhao,et al.  Generation of Targeted Point Mutations in Rice by a Modified CRISPR/Cas9 System. , 2017, Molecular plant.