Prospects for molecular farming in the green alga Chlamydomonas reinhardtii

Franklin, Scott; Mayfield, Stephen P.

doi:10.1016/j.pbi.2004.01.012

Cited by 119 publications

(56 citation statements)

References 38 publications

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“…3). It has been shown previously that C. reinhardtii chloroplast transformation results in very high transgenic expression levels in many cases (Franklin and Mayfield, 2004). However, the nuclear transformation of putative CrVTE2 was superior to chloroplast transformation of SynVTE2ct in an effort to obtain tocopherol-overproducing strains.…”

Section: Discussion Engineering Tocopherol Overproduction In C Reinhmentioning

confidence: 97%

Overlapping Photoprotective Function of Vitamin E and Carotenoids inChlamydomonas

Keasling

Niyogi

2011

Plant Physiology

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Tocopherols (vitamin E) and carotenoids are the two most abundant groups of lipid-soluble antioxidants in the chloroplast. Carotenoids are well known for their roles in protecting against photooxidative stress, whereas the photoprotective functions of tocopherols have only recently been examined experimentally. In addition, little is known about the functional overlap of carotenoids and tocopherols in vivo. To investigate this possible overlap, Chlamydomonas reinhardtii strains were engineered to overproduce tocopherols by chloroplast transformation with non-codon-optimized and codon-optimized versions of the homogentisate phytyltransferase vitamin E2 (VTE2) from Synechocystis and by nuclear transformation with VTE2 from C. reinhardtii, which resulted in 1.6-fold, 5-fold to 10-fold, and more than 10-fold increases in total tocopherol content, respectively. To test if tocopherol overproduction can compensate for carotenoid deficiency in terms of antioxidant function, the nuclear VTE2 gene from C. reinhardtii was overexpressed in the npq1 lor1 double mutant, which lacks zeaxanthin and lutein. Following transfer to high light, the npq1 lor1 strains that overaccumulated tocopherols showed increased resistance for up to 2 d and higher efficiency of photosystem II, and they were also much more resistant to other oxidative stresses. These results suggest an overlapping functions of tocopherols and carotenoids in protection against photooxidative stress.

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Section: Discussion Engineering Tocopherol Overproduction In C Reinhmentioning

confidence: 97%

Overlapping Photoprotective Function of Vitamin E and Carotenoids inChlamydomonas

Keasling

Niyogi

2011

Plant Physiology

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“…More recently we have shown that recombinant proteins also can be secreted from algae (8). Algae require only trace minerals, fertilizer, and sunlight to be grown at scale, giving them the potential to produce recombinant proteins, including therapeutics, very inexpensively (9). Although cost can be a significant factor in the production of protein-based therapies, producing unique classes of therapeutically relevant proteins is desirable, and algae offer the potential to produce a number of novel proteins because of the unique biochemical environment of the chloroplast (5).…”

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confidence: 99%

Production of unique immunotoxin cancer therapeutics in algal chloroplasts

Tran

Van

Barrera

et al. 2012

Proc. Natl. Acad. Sci. U.S.A.

165

118

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The idea of targeted therapy, whereby drug or protein molecules are delivered to specific cells, is a compelling approach to treating disease. Immunotoxins are one such targeted therapeutic, consisting of an antibody domain for binding target cells and molecules of a toxin that inhibits the proliferation of the targeted cell. One major hurdle preventing these therapies from reaching the market has been the lack of a suitable production platform that allows the cost-effective production of these highly complex molecules. The chloroplast of the green alga Chlamydomonas reinhardtii has been shown to contain the machinery necessary to fold and assemble complex eukaryotic proteins. However, the translational apparatus of chloroplasts resembles that of a prokaryote, allowing them to accumulate eukaryotic toxins that otherwise would kill a eukaryotic host. Here we show expression and accumulation of monomeric and dimeric immunotoxin proteins in algal chloroplasts. These fusion proteins contain an antibody domain targeting CD22, a B-cell surface epitope, and the enzymatic domain of exotoxin A from Pseudomonas aeruginosa. We demonstrated that algal-produced immunotoxins accumulate as soluble and enzymatically active proteins that bind target B cells and efficiently kill them in vitro. We also show that treatment with either the mono-or dimeric immunotoxins significantly prolongs the survival of mice with implanted human B-cell tumors.

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“…Интеграция фрагментов экзогенных ДНК в геном этой водоросли протекает по механизму гомологичной ре-комбинации. Клеточные популяции водорослей гомоген-ны по размеру клеток, клеточная масса удваивается через каждые 4-8 ч культивирования (Franklin, Mayfield, 2004). Для культивирования C. reinhardtii возможно использова-ние биореакторов больших объемов (до 500 тыс.…”

Section: актуальные технологии генетики и клеточной биологииunclassified

Plant expression systems for production of recombinant pharmaceutically important proteins

Дейнеко¹,

Загорская²

2017

Vestn. VOGiS

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The market of pharmaceutically valuable proteins is the fastest growing segment of the economy. Most biopharmaceuticals have been obtained in mammalian and microorganism cells, but both systems have a num ber of disadvantages. Plant cells combine the advan tages of the eukaryotic system of protein production and the simplicity and cheapness of the bacterial, and the use of plants for the production of recombinant proteins is an economically important and promising direction. The advantage of plant systems is the lower cost of cell cultivation. They are free from unwanted components, such as bacterial endotoxins, hyperglyco sylated proteins produced by yeast, animal and human pathogens in cell cultures of transgenic animals. In ad dition, plants are higher eukaryotes, and therefore full value folding and the formation of multimeric protein complexes occur in their cells, as well as a significant portion of posttranslational modifications similar to those in mammalian cells. The currently developed plant expression systems for recombinant proteins are extremely diverse and number more than 100 different technologies based on different plant species, gene transfer methods, expression strategies, methods for the subsequent extraction of the target protein, etc. This is nuclear and plastid transformation, transient and stable expression during transformation using agro bacterial transport, bombardment or electroporation, cultivation of whole terrestrial or aquatic plants, plant tissues or suspension cell cultures as expression sys tems. The review examines the current state of research in the use of plant expression systems for the produc tion of recombinant proteins for pharmaceuticals. The emphasis was placed on the advantages of plant cell cultures in comparison with other expression systems. Specific examples discuss promising plant systems for the production of recombinant proteins, such as trans plastomic plants, moss and aquatic plant cultures, as well as suspension cultures of cells of higher plants. The current state of the market for recombinant proteins obtained using plant expression systems is considered. The prospects of creating plant ("edible") vaccines based on genetically modified plants are discussed.Key words: expression systems; transgenic plants; bio producers; recombinant proteins; plant vaccines.Рынок фармацевтически ценных белков -наиболее быстро разви вающийся сегмент экономики. Большая часть биофармацевтиков получена в клетках млекопитающих и микроорганизмов, однако обе системы обладают рядом недостатков. Растительные клетки сочетают в себе достоинства эукариотической системы наработки белка и простоту и дешевизну бактериальной. Использование рас тений для получения рекомбинантных белков -экономически зна чимое и перспективное направление. Преимуществом раститель ных систем является более низкая стоимость культивирования клеток. Они свободны от нежелательных компонентов, таких как эндотоксины бактерий, гипергликозилированные белки, продуци руемые дрожжами, патогены животных и человека в кле...

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Prospects for molecular farming in the green alga Chlamydomonas reinhardtii

Cited by 119 publications

References 38 publications

Overlapping Photoprotective Function of Vitamin E and Carotenoids inChlamydomonas

Overlapping Photoprotective Function of Vitamin E and Carotenoids inChlamydomonas

Production of unique immunotoxin cancer therapeutics in algal chloroplasts

Plant expression systems for production of recombinant pharmaceutically important proteins

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