Creating and functionalising a synthetic compartment in plants

<p>Compartmentalisation is a universal feature of life, allowing organisms to separate their functions from each other and the environment. It enables greater control and complexity for cellular processes. Taking a cue from nature, engineered compartmentalisation has become one of the top prio...

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Main Author: Sandor, A
Other Authors: Sweetlove, L
Format: Thesis
Language:English
Published: 2022
Subjects:
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author Sandor, A
author2 Sweetlove, L
author_facet Sweetlove, L
Sandor, A
author_sort Sandor, A
collection OXFORD
description <p>Compartmentalisation is a universal feature of life, allowing organisms to separate their functions from each other and the environment. It enables greater control and complexity for cellular processes. Taking a cue from nature, engineered compartmentalisation has become one of the top priorities of synthetic biology. While many alternative strategies have emerged, building a synthetic membranous organelle de novo remains prohibitively difficult, despite its potential as a tool for metabolic engineering. This lack of an orthogonal synthetic organelle especially hampers the development of the nascent plant molecular farming industry. In this thesis, I describe the design and characterisation of a novel synthetic membranous compartment derived from the plant endoplasmic reticulum (ER), and proof-of-principle experiments to showcase its potential as a bioproduction platform in plants. The formation of the compartment is triggered by overexpression of a single recombinant scaffold protein, and I investigated the relationship between the structure of the scaffold protein and the morphology of the resultant compartment. I also compared the compartment to a conceptually similar ER rearrangement called organised smooth endoplasmic reticulum to determine their relation. Notably, the formation of the compartment had very little impact on the overall health of the host plant, and successful pilot experiments for the purification of the scaffold and other proteins anchored to it, as well as running enzymatic reactions on the surface of the compartment demonstrate the great potential of this system to serve as a plant synthetic biology tool. With further development, this platform could be a valuable asset to the plant biosynthesis and recombinant protein production industry.</p>
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spelling oxford-uuid:5d2002b3-142f-45ff-be10-dc160304d8e42022-07-28T10:23:29ZCreating and functionalising a synthetic compartment in plantsThesishttp://purl.org/coar/resource_type/c_db06uuid:5d2002b3-142f-45ff-be10-dc160304d8e4Genetic engineeringProtein engineeringEndoplasmic reticulumPlant organellesEnglishHyrax Deposit2022Sandor, ASweetlove, LFricker, MKirchhelle, CVan der hoorn, R<p>Compartmentalisation is a universal feature of life, allowing organisms to separate their functions from each other and the environment. It enables greater control and complexity for cellular processes. Taking a cue from nature, engineered compartmentalisation has become one of the top priorities of synthetic biology. While many alternative strategies have emerged, building a synthetic membranous organelle de novo remains prohibitively difficult, despite its potential as a tool for metabolic engineering. This lack of an orthogonal synthetic organelle especially hampers the development of the nascent plant molecular farming industry. In this thesis, I describe the design and characterisation of a novel synthetic membranous compartment derived from the plant endoplasmic reticulum (ER), and proof-of-principle experiments to showcase its potential as a bioproduction platform in plants. The formation of the compartment is triggered by overexpression of a single recombinant scaffold protein, and I investigated the relationship between the structure of the scaffold protein and the morphology of the resultant compartment. I also compared the compartment to a conceptually similar ER rearrangement called organised smooth endoplasmic reticulum to determine their relation. Notably, the formation of the compartment had very little impact on the overall health of the host plant, and successful pilot experiments for the purification of the scaffold and other proteins anchored to it, as well as running enzymatic reactions on the surface of the compartment demonstrate the great potential of this system to serve as a plant synthetic biology tool. With further development, this platform could be a valuable asset to the plant biosynthesis and recombinant protein production industry.</p>
spellingShingle Genetic engineering
Protein engineering
Endoplasmic reticulum
Plant organelles
Sandor, A
Creating and functionalising a synthetic compartment in plants
title Creating and functionalising a synthetic compartment in plants
title_full Creating and functionalising a synthetic compartment in plants
title_fullStr Creating and functionalising a synthetic compartment in plants
title_full_unstemmed Creating and functionalising a synthetic compartment in plants
title_short Creating and functionalising a synthetic compartment in plants
title_sort creating and functionalising a synthetic compartment in plants
topic Genetic engineering
Protein engineering
Endoplasmic reticulum
Plant organelles
work_keys_str_mv AT sandora creatingandfunctionalisingasyntheticcompartmentinplants