Plant Biotechnol J. 2021 Sep 29. doi: 10.1111/pbi.13717. Online ahead of print.
ABSTRACT
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 thirty-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: 1) episomal replication in plastids; 2) facile cloning; 3) predictable transgene expression in plastids; 4) non-integration of vector sequences into the endogenous plastome and; 5) 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.
PMID:34585834 | DOI:10.1111/pbi.13717