Site-specific recombinase genome engineering toolkit in maize

Jon P. Cody; Nathaniel D. Graham; Changzeng Zhao; Nathan C. Swyers; James A. Birchler

doi:10.1002/pld3.209

Site-specific recombinase genome engineering toolkit in maize

Jon P. Cody, Nathaniel D. Graham, Changzeng Zhao, Nathan C. Swyers, James A. Birchler

Genetics, Cell Biology and Development (CBS)

Research output: Contribution to journal › Article › peer-review

9 Scopus citations

Abstract

Site-specific recombinase enzymes function in heterologous cellular environments to initiate strand-switching reactions between unique DNA sequences termed recombinase binding sites. Depending on binding site position and orientation, reactions result in integrations, excisions, or inversions of targeted DNA sequences in a precise and predictable manner. Here, we established five different stable recombinase expression lines in maize through Agrobacterium-mediated transformation of T-DNA molecules that contain coding sequences for Cre, R, FLPe, phiC31 Integrase, and phiC31 excisionase. Through the bombardment of recombinase activated DsRed transient expression constructs, we have determined that all five recombinases are functional in maize plants. These recombinase expression lines could be utilized for a variety of genetic engineering applications, including selectable marker removal, targeted transgene integration into predetermined locations, and gene stacking.

Original language	English (US)
Article number	e00209
Journal	Plant Direct
Volume	4
Issue number	3
DOIs	https://doi.org/10.1002/pld3.209
State	Published - Mar 1 2020

Bibliographical note

Funding Information:
We thank Jan Schaart of Wageningen University & Research in Wageningen, Netherlands, for providing sequence information of R recombinase; Vibha Srivastava of the University of Arkansas for providing the FLPe coding sequence; Mario Gils of Leibniz-Institut fur Pflanzengenetik und Kulturpflanzenforschung (IPK), Gatersleben, Germany, for providing the phiC31 Integrase coding sequence; Michele Calos of Stanford University School of Medicine for providing phiC31 excisionase coding sequences; Stanton B. Gelvin of Purdue University for providing the Agrobacterium strains used in the transformation process; and the University of Missouri Molecular Cytology Core for providing support and access to stereo microscopy. Research on this topic was supported by a grant from the National Science Foundation, IOS-1339198.

Funding Information:
We thank Jan Schaart of Wageningen University & Research in Wageningen, Netherlands, for providing sequence information of R recombinase; Vibha Srivastava of the University of Arkansas for providing the FLPe coding sequence; Mario Gils of Leibniz‐Institut fur Pflanzengenetik und Kulturpflanzenforschung (IPK), Gatersleben, Germany, for providing the phiC31 Integrase coding sequence; Michele Calos of Stanford University School of Medicine for providing phiC31 excisionase coding sequences; Stanton B. Gelvin of Purdue University for providing the Agrobacterium strains used in the transformation process; and the University of Missouri Molecular Cytology Core for providing support and access to stereo microscopy. Research on this topic was supported by a grant from the National Science Foundation, IOS‐1339198.

Publisher Copyright:
© 2020 The Authors. Plant Direct published by American Society of Plant Biologists, Society for Experimental Biology and John Wiley & Sons Ltd.

Keywords

Agrobacterium
bombardment
genetic engineering
maize
recombinases

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10.1002/pld3.209

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title = "Site-specific recombinase genome engineering toolkit in maize",

abstract = "Site-specific recombinase enzymes function in heterologous cellular environments to initiate strand-switching reactions between unique DNA sequences termed recombinase binding sites. Depending on binding site position and orientation, reactions result in integrations, excisions, or inversions of targeted DNA sequences in a precise and predictable manner. Here, we established five different stable recombinase expression lines in maize through Agrobacterium-mediated transformation of T-DNA molecules that contain coding sequences for Cre, R, FLPe, phiC31 Integrase, and phiC31 excisionase. Through the bombardment of recombinase activated DsRed transient expression constructs, we have determined that all five recombinases are functional in maize plants. These recombinase expression lines could be utilized for a variety of genetic engineering applications, including selectable marker removal, targeted transgene integration into predetermined locations, and gene stacking.",

keywords = "Agrobacterium, bombardment, genetic engineering, maize, recombinases",

author = "Cody, {Jon P.} and Graham, {Nathaniel D.} and Changzeng Zhao and Swyers, {Nathan C.} and Birchler, {James A.}",

note = "Funding Information: We thank Jan Schaart of Wageningen University & Research in Wageningen, Netherlands, for providing sequence information of R recombinase; Vibha Srivastava of the University of Arkansas for providing the FLPe coding sequence; Mario Gils of Leibniz-Institut fur Pflanzengenetik und Kulturpflanzenforschung (IPK), Gatersleben, Germany, for providing the phiC31 Integrase coding sequence; Michele Calos of Stanford University School of Medicine for providing phiC31 excisionase coding sequences; Stanton B. Gelvin of Purdue University for providing the Agrobacterium strains used in the transformation process; and the University of Missouri Molecular Cytology Core for providing support and access to stereo microscopy. Research on this topic was supported by a grant from the National Science Foundation, IOS-1339198. Funding Information: We thank Jan Schaart of Wageningen University & Research in Wageningen, Netherlands, for providing sequence information of R recombinase; Vibha Srivastava of the University of Arkansas for providing the FLPe coding sequence; Mario Gils of Leibniz‐Institut fur Pflanzengenetik und Kulturpflanzenforschung (IPK), Gatersleben, Germany, for providing the phiC31 Integrase coding sequence; Michele Calos of Stanford University School of Medicine for providing phiC31 excisionase coding sequences; Stanton B. Gelvin of Purdue University for providing the Agrobacterium strains used in the transformation process; and the University of Missouri Molecular Cytology Core for providing support and access to stereo microscopy. Research on this topic was supported by a grant from the National Science Foundation, IOS‐1339198. Publisher Copyright: {\textcopyright} 2020 The Authors. Plant Direct published by American Society of Plant Biologists, Society for Experimental Biology and John Wiley & Sons Ltd.",

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doi = "10.1002/pld3.209",

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AU - Graham, Nathaniel D.

AU - Zhao, Changzeng

AU - Swyers, Nathan C.

AU - Birchler, James A.

N1 - Funding Information: We thank Jan Schaart of Wageningen University & Research in Wageningen, Netherlands, for providing sequence information of R recombinase; Vibha Srivastava of the University of Arkansas for providing the FLPe coding sequence; Mario Gils of Leibniz-Institut fur Pflanzengenetik und Kulturpflanzenforschung (IPK), Gatersleben, Germany, for providing the phiC31 Integrase coding sequence; Michele Calos of Stanford University School of Medicine for providing phiC31 excisionase coding sequences; Stanton B. Gelvin of Purdue University for providing the Agrobacterium strains used in the transformation process; and the University of Missouri Molecular Cytology Core for providing support and access to stereo microscopy. Research on this topic was supported by a grant from the National Science Foundation, IOS-1339198. Funding Information: We thank Jan Schaart of Wageningen University & Research in Wageningen, Netherlands, for providing sequence information of R recombinase; Vibha Srivastava of the University of Arkansas for providing the FLPe coding sequence; Mario Gils of Leibniz‐Institut fur Pflanzengenetik und Kulturpflanzenforschung (IPK), Gatersleben, Germany, for providing the phiC31 Integrase coding sequence; Michele Calos of Stanford University School of Medicine for providing phiC31 excisionase coding sequences; Stanton B. Gelvin of Purdue University for providing the Agrobacterium strains used in the transformation process; and the University of Missouri Molecular Cytology Core for providing support and access to stereo microscopy. Research on this topic was supported by a grant from the National Science Foundation, IOS‐1339198. Publisher Copyright: © 2020 The Authors. Plant Direct published by American Society of Plant Biologists, Society for Experimental Biology and John Wiley & Sons Ltd.

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N2 - Site-specific recombinase enzymes function in heterologous cellular environments to initiate strand-switching reactions between unique DNA sequences termed recombinase binding sites. Depending on binding site position and orientation, reactions result in integrations, excisions, or inversions of targeted DNA sequences in a precise and predictable manner. Here, we established five different stable recombinase expression lines in maize through Agrobacterium-mediated transformation of T-DNA molecules that contain coding sequences for Cre, R, FLPe, phiC31 Integrase, and phiC31 excisionase. Through the bombardment of recombinase activated DsRed transient expression constructs, we have determined that all five recombinases are functional in maize plants. These recombinase expression lines could be utilized for a variety of genetic engineering applications, including selectable marker removal, targeted transgene integration into predetermined locations, and gene stacking.

AB - Site-specific recombinase enzymes function in heterologous cellular environments to initiate strand-switching reactions between unique DNA sequences termed recombinase binding sites. Depending on binding site position and orientation, reactions result in integrations, excisions, or inversions of targeted DNA sequences in a precise and predictable manner. Here, we established five different stable recombinase expression lines in maize through Agrobacterium-mediated transformation of T-DNA molecules that contain coding sequences for Cre, R, FLPe, phiC31 Integrase, and phiC31 excisionase. Through the bombardment of recombinase activated DsRed transient expression constructs, we have determined that all five recombinases are functional in maize plants. These recombinase expression lines could be utilized for a variety of genetic engineering applications, including selectable marker removal, targeted transgene integration into predetermined locations, and gene stacking.

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Site-specific recombinase genome engineering toolkit in maize

Abstract

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