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Genetic Engineering of Microalgae
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Genetic Engineering of Microalgae

A special issue of Genes (ISSN 2073-4425). This special issue belongs to the section "Microbial Genetics and Genomics".

Deadline for manuscript submissions: closed (20 January 2023) | Viewed by 6383

Special Issue Editor

Prof. Dr. Armin Hallmann

E-Mail Website
Guest Editor
Department of Cellular and Developmental Biology of Plants, University of Bielefeld, 33615 Bielefeld, Germany
Interests: microalgae; microbial genetics; molecular biology; algal transgenics and biotechnology; genetic engineering; genetics of cell differentiation and multicellularity; green algae

Special Issue Information

Dear Colleagues,

Microalgae provide a wide variety of biomolecules, cellular functions, and physiological characteristics not least due to the great species diversity and the different living conditions within this polyphyletic group of photosynthetic microorganisms. These and other reasons mean that microalgae are common target organisms for genetic modification, ranging from basic research to commercial biotechnological applications. Recent developments in genetic engineering of microalgae have been supported by rapid advances in omics technologies and an increase in methods and tools for gene manipulation, vector construction, transformation, and selection. Even target-specific in vivo modification of the genome has been made possible by chimeric genome-editing tools.

This Special Issue welcomes both reviews and original articles on the genetic engineering of microalgae. Possible contents include but are not limited to systems biology and omics technologies in relation to genetic engineering of microalgae (e.g., genomics, transcriptomics, proteomics, metabolomics), key tools or elements of genetic engineering (e.g., gene or metabolic pathway design, selectable marker and reporter genes, regulatory elements, transformation techniques, gene silencing, gene knockout, genome editing) and the application of genetic engineering in basic research and industry (e.g., biofuel production, secondary metabolite production, optogenetics).

We eagerly await your contributions.

Prof. Dr. Armin Hallmann
Guest Editor

Manuscript Submission Information

Manuscripts should be submitted online at www.mdpi.com by registering and logging in to this website. Once you are registered, click here to go to the submission form. Manuscripts can be submitted until the deadline. All submissions that pass pre-check are peer-reviewed. Accepted papers will be published continuously in the journal (as soon as accepted) and will be listed together on the special issue website. Research articles, review articles as well as short communications are invited. For planned papers, a title and short abstract (about 100 words) can be sent to the Editorial Office for announcement on this website.

Submitted manuscripts should not have been published previously, nor be under consideration for publication elsewhere (except conference proceedings papers). All manuscripts are thoroughly refereed through a single-blind peer-review process. A guide for authors and other relevant information for submission of manuscripts is available on the Instructions for Authors page. Genes is an international peer-reviewed open access monthly journal published by MDPI.

Please visit the Instructions for Authors page before submitting a manuscript. The Article Processing Charge (APC) for publication in this open access journal is 2600 CHF (Swiss Francs). Submitted papers should be well formatted and use good English. Authors may use MDPI's English editing service prior to publication or during author revisions.

Keywords

  • microalgae
  • omics
  • algal transformation
  • transgenic algae
  • genetic engineering
  • genetic modification
  • forward genetics
  • reverse genetics
  • genome-editing tools
  • synthetic biology
  • bioengineering
  • transgene expression
  • biotechnological production
  • algal cell factories
  • production of lipids and carotenoids
  • secondary metabolite production

Published Papers (3 papers)

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20 pages, 5592 KiB  
Article
Cell Type-Specific Promoters of Volvox carteri for Molecular Cell Biology Studies
by Benjamin von der Heyde, Eva Laura von der Heyde and Armin Hallmann
Genes 2023, 14(7), 1389; https://doi.org/10.3390/genes14071389 - 01 Jul 2023
Viewed by 2418
Abstract
The multicellular green alga Volvox carteri has emerged as a valuable model organism for investigating various aspects of multicellularity and cellular differentiation, photoreception and phototaxis, cell division, biogenesis of the extracellular matrix and morphogenetic movements. While a range of molecular tools and bioinformatics [...] Read more.
The multicellular green alga Volvox carteri has emerged as a valuable model organism for investigating various aspects of multicellularity and cellular differentiation, photoreception and phototaxis, cell division, biogenesis of the extracellular matrix and morphogenetic movements. While a range of molecular tools and bioinformatics resources have been made available for exploring these topics, the establishment of cell type-specific promoters in V. carteri has not been achieved so far. Therefore, here, we conducted a thorough screening of transcriptome data from RNA sequencing analyses of V. carteri in order to identify potential cell type-specific promoters. Eventually, we chose two putative strong and cell type-specific promoters, with one exhibiting specific expression in reproductive cells (gonidia), the PCY1 promoter, and the other in somatic cells, the PFP promoter. After cloning both promoter regions, they were introduced upstream of a luciferase reporter gene. By using particle bombardment, the DNA constructs were stably integrated into the genome of V. carteri. The results of the expression analyses, which were conducted at both the transcript and protein levels, demonstrated that the two promoters drive cell type-specific expression in their respective target cell types. Transformants with considerably diverse expression levels of the chimeric genes were identifiable. In conclusion, the screening and analysis of transcriptome data from RNA sequencing allowed for the identification of potential cell type-specific promoters in V. carteri. Reporter gene constructs demonstrated the actual usability of two promoters. The investigated PCY1 and PFP promoters were proven to be potent molecular tools for genetic engineering in V. carteri. Full article
(This article belongs to the Special Issue Genetic Engineering of Microalgae)
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15 pages, 2031 KiB  
Article
New Synthetic Operon Vectors for Expressing Multiple Proteins in the Chlamydomonas reinhardtii Chloroplast
by Jihye Yeon, Stephen M. Miller and Wipawee Dejtisakdi
Genes 2023, 14(2), 368; https://doi.org/10.3390/genes14020368 - 31 Jan 2023
Cited by 2 | Viewed by 1793
Abstract
Microalgae are a promising platform for generating valuable commercial products, including proteins that may not express well in more traditional cell culture systems. In the model green alga Chlamydomonas reinhardtii, transgenic proteins can be expressed from either the nuclear or chloroplast genome. [...] Read more.
Microalgae are a promising platform for generating valuable commercial products, including proteins that may not express well in more traditional cell culture systems. In the model green alga Chlamydomonas reinhardtii, transgenic proteins can be expressed from either the nuclear or chloroplast genome. Expression in the chloroplast has several advantages, but technology is not yet well developed for expressing multiple transgenic proteins simultaneously. Here, we developed new synthetic operon vectors to express multiple proteins from a single chloroplast transcription unit. We modified an existing chloroplast expression vector to contain intercistronic elements derived from cyanobacterial and tobacco operons and tested the ability of the resulting operon vectors to express two or three different proteins at a time. All operons containing two of the coding sequences (for C. reinhardtii FBP1 and atpB) expressed the products of those genes, but operons containing the other two coding sequences (C. reinhardtii FBA1 and the synthetic camelid antibody gene VHH) did not. These results expand the repertoire of intercistronic spacers that can function in the C. reinhardtii chloroplast, but they also suggest that some coding sequences do not function well in the context of synthetic operons in this alga. Full article
(This article belongs to the Special Issue Genetic Engineering of Microalgae)
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11 pages, 2459 KiB  
Brief Report
Analysis of Viral Promoters for Transgene Expression and of the Effect of 5′-UTRs on Alternative Translational Start Sites in Chlamydomonas
by Justus Niemeyer, Laura Fischer, Frank O’Neill Aylward and Michael Schroda
Genes 2023, 14(4), 948; https://doi.org/10.3390/genes14040948 - 21 Apr 2023
Cited by 1 | Viewed by 1817
Abstract
Microalgae biotechnology has the potential to produce high quality bioproducts in a sustainable manner. Here, Chlamydomonas reinhardtii has shown great potential as a host for biotechnological exploitation. However, low expression of nuclear transgenes is still a problem and needs to be optimized. In [...] Read more.
Microalgae biotechnology has the potential to produce high quality bioproducts in a sustainable manner. Here, Chlamydomonas reinhardtii has shown great potential as a host for biotechnological exploitation. However, low expression of nuclear transgenes is still a problem and needs to be optimized. In many model organisms, viral promoters are used to drive transgene expression at high levels. However, no viruses are known to infect Chlamydomonas, and known viral promoters are not functional. Recently, two different lineages of giant viruses were identified in the genomes of Chlamydomonas reinhardtii field isolates. In this work, we tested six potentially strong promoters from these viral genomes for their ability to drive transgene expression in Chlamydomonas. We used ble, NanoLUC, and mCherry as reporter genes, and three native benchmark promoters as controls. None of the viral promoters drove expression of any reporter gene beyond background. During our study, we found that mCherry variants are produced by alternative in-frame translational start sites in Chlamydomonas. We show that this problem can be overcome by mutating the responsible methionine codons to codons for leucine and by using the 5′-UTR of βTUB2 instead of the 5′-UTRs of PSAD or RBCS2. Apparently, the βTUB2 5′-UTR promotes the use of the first start codon. This could be mediated by the formation of a stem-loop between sequences of the βTUB2 5′-UTR and sequences downstream of the first AUG in the mCherry reporter, potentially increasing the dwell time of the scanning 40S subunit on the first AUG and thus decreasing the probability of leaky scanning. Full article
(This article belongs to the Special Issue Genetic Engineering of Microalgae)
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