Expression of an HBcAg-based antigen carrying angiotensin II in Chlamydomonas reinhardtii as a candidate hypertension vaccine

Soria-Guerra, Ruth Elena; Ramírez-Alonso, Jocelin Itzel; Ibáñez-Salazar, Alejandro; Govea‐Alonso, Dania O.; Paz-Maldonado, Luz M. T.; Bañuelos-Hernández, Bernardo; Korban, Schuyler S.; Rosales‐Mendoza, Sergio

doi:10.1007/s11240-013-0388-x

Cited by 30 publications

(12 citation statements)

References 24 publications

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“…Many heterologous transgenes have been expressed in Chlamydomonas , including many antibiotic resistance markers, reporter genes, and commercially relevant proteins, such as antigens for vaccines (Soria‐Guerra et al ., ).…”

Section: Expression Of Transgenes In the Chlamydomonas Nuclear Genomementioning

confidence: 97%

“…The expression of transgenes in the nuclear genome of Chlamydomonas has many basic and applied applications Many heterologous transgenes have been expressed in Chlamydomonas, including many antibiotic resistance markers, reporter genes, and commercially relevant proteins, such as antigens for vaccines (Soria-Guerra et al, 2013).…”

Section: Expression Of Transgenes In the Chlamydomonas Nuclear Genomementioning

confidence: 99%

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Molecular techniques to interrogate and edit the Chlamydomonas nuclear genome

2015

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SUMMARYThe success of the green alga Chlamydomonas reinhardtii as a model organism is to a large extent due to the wide range of molecular techniques that are available for its characterization. Here, we review some of the techniques currently used to modify and interrogate the C. reinhardtii nuclear genome and explore several technologies under development. Nuclear mutants can be generated with ultraviolet (UV) light and chemical mutagens, or by insertional mutagenesis. Nuclear transformation methods include biolistic delivery, agitation with glass beads, and electroporation. Transforming DNA integrates into the genome at random sites, and multiple strategies exist for mapping insertion sites. A limited number of studies have demonstrated targeted modification of the nuclear genome by approaches such as zinc-finger nucleases and homologous recombination. RNA interference is widely used to knock down expression levels of nuclear genes. A wide assortment of transgenes has been successfully expressed in the Chlamydomonas nuclear genome, including transformation markers, fluorescent proteins, reporter genes, epitope tagged proteins, and even therapeutic proteins. Optimized expression constructs and strains help transgene expression. Emerging technologies such as the CRISPR/Cas9 system, high-throughput mutant identification, and a whole-genome knockout library are being developed for this organism. We discuss how these advances will propel future investigations.

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Section: Expression Of Transgenes In the Chlamydomonas Nuclear Genomementioning

confidence: 97%

Section: Expression Of Transgenes In the Chlamydomonas Nuclear Genomementioning

confidence: 99%

Molecular techniques to interrogate and edit the Chlamydomonas nuclear genome

2015

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“…A chimeric protein, angiotensin II fused with a nucleocapsid antigen of hepatitis B virus (HBcAg) was produced in C. reinhardtii . The expression levels of this fused protein reached 0.05% of TSP in several transgenic lines; but the immunogenic properties of antigen derived from the HBcAgII algae still need to be assessed [ 176 ].…”

Section: Genetic Engineering and Production Of Pharmaceuticals Andmentioning

confidence: 99%

The Potential for Microalgae as Bioreactors to Produce Pharmaceuticals

Yan

Fan

Chen

et al. 2016

IJMS

178

102

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As photosynthetic organisms, microalgae can efficiently convert solar energy into biomass. Microalgae are currently used as an important source of valuable natural biologically active molecules, such as carotenoids, chlorophyll, long-chain polyunsaturated fatty acids, phycobiliproteins, carotenoids and enzymes. Significant advances have been achieved in microalgae biotechnology over the last decade, and the use of microalgae as bioreactors for expressing recombinant proteins is receiving increased interest. Compared with the bioreactor systems that are currently in use, microalgae may be an attractive alternative for the production of pharmaceuticals, recombinant proteins and other valuable products. Products synthesized via the genetic engineering of microalgae include vaccines, antibodies, enzymes, blood-clotting factors, immune regulators, growth factors, hormones, and other valuable products, such as the anticancer agent Taxol. In this paper, we briefly compare the currently used bioreactor systems, summarize the progress in genetic engineering of microalgae, and discuss the potential for microalgae as bioreactors to produce pharmaceuticals.

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“…The nuclear genome of C. reinhardtii has been sequenced [ 40 ] and many mutants have been created for molecular biology research or heterologous expression of useful proteins. To date, a lot of functional enzymes [ 41 ], antigens [ 42 ], antibodies [ 43 ], and vaccines [ 44 ] have been successfully expressed in C. reinhardtii . As a platform for protein production, C. reinhardtii has certain advantages, including rapid growth, capability of protein post-translational processing, and expressing a large amount of foreign protein in chloroplasts.…”

Section: Introductionmentioning

confidence: 99%

Expression of Anti-Lipopolysaccharide Factor Isoform 3 in Chlamydomonas reinhardtii Showing High Antimicrobial Activity

Huang

Wang

et al. 2021

Marine Drugs

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Antimicrobial peptides are a class of proteins with antibacterial functions. In this study, the anti-lipopolysaccharide factor isoform 3 gene (ALFPm3), encoding an antimicrobial peptide from Penaeus monodon with a super activity was expressed in Chlamydomonas reinhardtii, which would develop a microalga strain that can be used for the antimicrobial peptide production. To construct the expression cluster, namely pH2A-Pm3, the codon optimized ALFPm3 gene was fused with the ble reporter by 2A peptide and inserted into pH124 vector. The glass-bead method was performed to transform pH2A-Pm3 into C. reinhardtii CC-849. In addition to 8 μg/mL zeocin resistance selection, the C. reinhardtii transformants were further confirmed by genomic PCR and RT-PCR. Western blot analysis showed that the C. reinhardtii-derived ALFPm3 (cALFPm3) was successfully expressed in C. reinhardtii transformants and accounted for 0.35% of the total soluble protein (TSP). Furthermore, the results of antibacterial assay revealed that the cALFPm3 could significantly inhibit the growth of a variety of bacteria, including both Gram-negative bacteria and Gram-positive bacteria at a concentration of 0.77 μM. Especially, the inhibition could last longer than 24 h, which performed better than ampicillin. Hence, this study successfully developed a transgenic C. reinhardtii strain, which can produce the active ALFPm3 driven from P. monodon, providing a potential strategy to use C. reinhardtii as the cell factory to produce antimicrobial peptides.

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Expression of an HBcAg-based antigen carrying angiotensin II in Chlamydomonas reinhardtii as a candidate hypertension vaccine

Cited by 30 publications

References 24 publications

Molecular techniques to interrogate and edit the Chlamydomonas nuclear genome

Molecular techniques to interrogate and edit the Chlamydomonas nuclear genome

The Potential for Microalgae as Bioreactors to Produce Pharmaceuticals

Expression of Anti-Lipopolysaccharide Factor Isoform 3 in Chlamydomonas reinhardtii Showing High Antimicrobial Activity

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