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Title: Mini‐synplastomes for plastid genetic engineering

Abstract

Summary In the age of synthetic biology, plastid engineering requires a nimble platform to introduce novel synthetic circuits in plants. While effective for integrating relatively small constructs into the plastome, plastid engineering via homologous recombination of transgenes is over 30 years old. Here we show the design–build–test of a novel synthetic genome structure that does not disturb the native plastome: the ‘mini‐synplastome’. The mini‐synplastome was inspired by dinoflagellate plastome organization, which is comprised of numerous minicircles residing in the plastid instead of a single organellar genome molecule. The first mini‐synplastome in plants was developed in vitro to meet the following criteria: (i) episomal replication in plastids; (ii) facile cloning; (iii) predictable transgene expression in plastids; (iv) non‐integration of vector sequences into the endogenous plastome; and (v) autonomous persistence in the plant over generations in the absence of exogenous selection pressure. Mini‐synplastomes are anticipated to revolutionize chloroplast biotechnology, enable facile marker‐free plastid engineering, and provide an unparalleled platform for one‐step metabolic engineering in plants.

Authors:
 [1];  [1];  [1];  [2];  [2];  [1];  [3];  [3]; ORCiD logo [4]; ORCiD logo [2]; ORCiD logo [1]
  1. Department of Food Science University of Tennessee Knoxville TN USA, Center for Agricultural Synthetic Biology University of Tennessee Institute of Agriculture Knoxville TN USA
  2. Center for Agricultural Synthetic Biology University of Tennessee Institute of Agriculture Knoxville TN USA, Department of Plant Sciences University of Tennessee Knoxville TN USA
  3. Department of Plant Sciences University of Tennessee Knoxville TN USA
  4. Department of Basic and Translational Sciences School of Dental Medicine University of Pennsylvania Philadelphia PA USA
Publication Date:
Sponsoring Org.:
USDOE
OSTI Identifier:
1827240
Alternate Identifier(s):
OSTI ID: 1827242
Resource Type:
Published Article
Journal Name:
Plant Biotechnology Journal
Additional Journal Information:
Journal Name: Plant Biotechnology Journal Journal Volume: 20 Journal Issue: 2; Journal ID: ISSN 1467-7644
Publisher:
Wiley-Blackwell
Country of Publication:
United Kingdom
Language:
English

Citation Formats

Occhialini, Alessandro, Pfotenhauer, Alexander C., Li, Li, Harbison, Stacee A., Lail, Andrew J., Burris, Jason N., Piasecki, Cristiano, Piatek, Agnieszka A., Daniell, Henry, Stewart, Jr, C. Neal, and Lenaghan, Scott C.. Mini‐synplastomes for plastid genetic engineering. United Kingdom: N. p., 2021. Web. doi:10.1111/pbi.13717.
Occhialini, Alessandro, Pfotenhauer, Alexander C., Li, Li, Harbison, Stacee A., Lail, Andrew J., Burris, Jason N., Piasecki, Cristiano, Piatek, Agnieszka A., Daniell, Henry, Stewart, Jr, C. Neal, & Lenaghan, Scott C.. Mini‐synplastomes for plastid genetic engineering. United Kingdom. https://doi.org/10.1111/pbi.13717
Occhialini, Alessandro, Pfotenhauer, Alexander C., Li, Li, Harbison, Stacee A., Lail, Andrew J., Burris, Jason N., Piasecki, Cristiano, Piatek, Agnieszka A., Daniell, Henry, Stewart, Jr, C. Neal, and Lenaghan, Scott C.. Sun . "Mini‐synplastomes for plastid genetic engineering". United Kingdom. https://doi.org/10.1111/pbi.13717.
@article{osti_1827240,
title = {Mini‐synplastomes for plastid genetic engineering},
author = {Occhialini, Alessandro and Pfotenhauer, Alexander C. and Li, Li and Harbison, Stacee A. and Lail, Andrew J. and Burris, Jason N. and Piasecki, Cristiano and Piatek, Agnieszka A. and Daniell, Henry and Stewart, Jr, C. Neal and Lenaghan, Scott C.},
abstractNote = {Summary In the age of synthetic biology, plastid engineering requires a nimble platform to introduce novel synthetic circuits in plants. While effective for integrating relatively small constructs into the plastome, plastid engineering via homologous recombination of transgenes is over 30 years old. Here we show the design–build–test of a novel synthetic genome structure that does not disturb the native plastome: the ‘mini‐synplastome’. The mini‐synplastome was inspired by dinoflagellate plastome organization, which is comprised of numerous minicircles residing in the plastid instead of a single organellar genome molecule. The first mini‐synplastome in plants was developed in vitro to meet the following criteria: (i) episomal replication in plastids; (ii) facile cloning; (iii) predictable transgene expression in plastids; (iv) non‐integration of vector sequences into the endogenous plastome; and (v) autonomous persistence in the plant over generations in the absence of exogenous selection pressure. Mini‐synplastomes are anticipated to revolutionize chloroplast biotechnology, enable facile marker‐free plastid engineering, and provide an unparalleled platform for one‐step metabolic engineering in plants.},
doi = {10.1111/pbi.13717},
journal = {Plant Biotechnology Journal},
number = 2,
volume = 20,
place = {United Kingdom},
year = {2021},
month = {10}
}

Journal Article:
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https://doi.org/10.1111/pbi.13717

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