Open AccessMini‐synplastomes for plastid genetic engineering
Author(s) -
Occhialini Alessandro,
Pfotenhauer Alexander C.,
Li Li,
Harbison Stacee A.,
Lail Andrew J.,
Burris Jason N.,
Piasecki Cristiano,
Piatek Agnieszka A.,
Daniell Henry,
Stewart C. Neal,
Lenaghan Scott C.
Publication year - 2022
Publication title -
plant biotechnology journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.525
H-Index - 115
eISSN - 1467-7652
pISSN - 1467-7644
DOI - 10.1111/pbi.13717
Subject(s) - plastid , biology , genome engineering , minicircle , chloroplast dna , synthetic biology , genome , chloroplast , transgene , homologous recombination , computational biology , genetics , dna , gene , genome editing
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.