Abstract
Chlamydomonas reinhardtii is a well-established model system for basic research questions ranging from photosynthesis and organelle biogenesis, to the biology of cilia and basal bodies, to channelrhodopsins and photoreceptors. More recently, Chlamydomonas has also been recognized as a suitable host for the production of high-value chemicals and high-value recombinant proteins. However, basic and applied research have suffered from the inefficient expression of nuclear transgenes. The combined efforts of the Chlamydomonas community over the past decades have provided insights into the mechanisms underlying this phenomenon and have resulted in mutant strains defective in some silencing mechanisms. Moreover, many insights have been gained into the parameters that affect nuclear transgene expression, like promoters, introns, codon usage, or terminators. Here I critically review these insights and try to integrate them into design suggestions for the construction of nuclear transgenes that are to be expressed at high levels.
Highlights
Chlamydomonas reinhardtii is a well-established model system for basic research questions ranging from photosynthesis and organelle biogenesis, to the biology of cilia and basal bodies, to channelrhodopsins and photoreceptors
Chlamydomonas has emerged as a valuable model organism for basic research e.g., on photosynthesis and chloroplast biogenesis [2], the biology of cilia and basal bodies [3], the cell cycle [4], the plant heat stress response [5], or the circadian clock [6]
I provide an overview to the history of nuclear transformation and the pitfalls encountered regarding the expression of nuclear transgenes
Summary
Chlamydomonas reinhardtii is a unicellular green microalga living in the soil and in the pelagic zone of lakes [1]. Chlamydomonas has been recognized as a host for the production of high-value chemicals and high-value recombinant proteins [14,15]. For the latter, the Chlamydomonas chloroplast is a suitable expression platform with yields reported to range between 0.5% and 5% of total soluble protein [16]. Transgenes expressed in the nucleus, can be targeted for secretion, allowing recombinant proteins to be glycosylated and secreted into the medium, from where purification can be achieved more [17,18]. The major limitation of nuclear transgene expression in Chlamydomonas has been the low level of expression achieved. I try to integrate the insights gained into design suggestions for the construction of nuclear transgenes that are to be expressed at high levels
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