Abstract Microalgae chloroplasts are an attractive platform for the production of recombinant proteins, and are targets of considerable research aiming to optimise carbon fixation and lipid biosynthesis. Despite this, the potential of chloroplast transformation is currently underexplored, due to it being time-consuming and costly. It is further hindered by the lack of standardised genetic parts available. We are developing biological methods and open-source hardware to remove these bottlenecks and building a library of modular genetic components for Chlamydomonas chloroplast. Content Introduction Overview of biology part Overview of hardware part Hardware specifications Questions to labs / community groups A. Introduction: The traditional approach to production of recombinant proteins in bacteria is significantly limited, because bacteria lack most post-translational modifications, and are likely to produce proteins as insoluble aggregates. On the other hand, mammalian expression systems have poor expression yields, require complex media and growth conditions, and are very sensitive to contamination with pathogens. The use of microalgae chloroplasts as an alternative expression system solves most of these issues and offers many other advantages. Microalgae chloroplasts have very efficient protein expression. They also have a range of post-translational modification pathways, and possess chaperones and disulfide isomerases, which allow for the correct folding of proteins. Unlike in higher plants, metabolic energy in microalgae is not directed towards maintaining complex differentiated structures, further enhancing transgene expression (which can total up to 30-50% of a dry biomass). cell’s In addition to the outstanding transgene yields, microalgae are also very amenable to farming worldwide and outside of labs, as autotrophic, heterotrophic and mixotrophic growing conditions
Abstract Microalgae chloroplasts are an attractive platform for the production of recombinant proteins, and are targets of considerable research aiming to optimise carbon fixation and lipid biosynthesis. Despite this, the potential of chloroplast transformation is currently underexplored, due to it being time-consuming and costly. It is further hindered by the lack of standardised genetic parts available. We are developing biological methods and open-source hardware to remove these bottlenecks and building a library of modular genetic components for Chlamydomonas chloroplast. Content Introduction Overview of biology part Overview of hardware part Hardware specifications Questions to labs / community groups A. Introduction: The traditional approach to production of recombinant proteins in bacteria is significantly limited, because bacteria lack most post-translational modifications, and are likely to produce proteins as insoluble aggregates. On the other hand, mammalian expression systems have poor expression yields, require complex media and growth conditions, and are very sensitive to contamination with pathogens. The use of microalgae chloroplasts as an alternative expression system solves most of these issues and offers many other advantages. Microalgae chloroplasts have very efficient protein expression. They also have a range of post-translational modification pathways, and possess chaperones and disulfide isomerases, which allow for the correct folding of proteins. Unlike in higher plants, metabolic energy in microalgae is not directed towards maintaining complex differentiated structures, further enhancing transgene expression (which can total up to 30-50% of a cell’s dry biomass). In addition to the outstanding transgene yields, microalgae are also very amenable to farming worldwide and outside of labs, as autotrophic, heterotrophic and mixotrophic growing conditions
T--Cambridge-JIC--HP--Questions.docx