Rational Design and Screening of Synthetic Promoters in Chlamydomonas reinhardtii.
Chlamydomonas reinhardtii
Genetic transformation
Synthetic biology
Synthetic promoters
Targeted mutagenesis
cis-regulatory elements
Journal
Methods in molecular biology (Clifton, N.J.)
ISSN: 1940-6029
Titre abrégé: Methods Mol Biol
Pays: United States
ID NLM: 9214969
Informations de publication
Date de publication:
2024
2024
Historique:
medline:
28
7
2024
pubmed:
28
7
2024
entrez:
27
7
2024
Statut:
ppublish
Résumé
Synthetic promoters are powerful tools to boost the biotechnological potential of microalgae as eco-sustainable industrial hosts. The increasing availability of transcriptome data on microalgae in a variety of environmental conditions allows to identify cis-regulatory elements (CREs) that are responsible for the transcriptional output. Furthermore, advanced cloning technologies, such as golden gate-based MoClo toolkits, enable the creation of modular constructs for testing multiple promoters and a range of reporter systems in a convenient manner. In this chapter, we will describe how to introduce in silico-identified CREs into promoter sequences, and how to clone the modified promoters into MoClo compatible vectors. We will then describe how these promoters can be evaluated and screened for transgene expression in an established microalgal model for genetic perturbation, i.e., Chlamydomonas reinhardtii.
Identifiants
pubmed: 39068332
doi: 10.1007/978-1-0716-4063-0_4
doi:
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
69-83Informations de copyright
© 2024. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.
Références
Milito A, Aschern M, McQuillan JL, Yang JS (2023) Challenges and advances towards the rational design of microalgal synthetic promoters in Chlamydomonas reinhardtii. J Exp Bot 74:3833–3850
doi: 10.1093/jxb/erad100
pubmed: 37025006
Venter M (2007) Synthetic promoters: genetic control through cis engineering. Trends Plant Sci 12:118–124
doi: 10.1016/j.tplants.2007.01.002
pubmed: 17292658
Mori Y, Shirai T (2018) Designing artificial metabolic pathways, construction of target enzymes, and analysis of their function. Curr Opin Biotechnol 54:41–44
doi: 10.1016/j.copbio.2018.01.021
pubmed: 29452926
Tas H, Grozinger L, Stoof R, de Lorenzo V, Goñi-Moreno A (2021) Contextual dependencies expand the re-usability of genetic inverters. Nat Commun 12:355
doi: 10.1038/s41467-020-20656-5
pubmed: 33441561
pmcid: 7806840
Filo M, Kumar S, Khammash M (2022) A hierarchy of biomolecular proportional-integral-derivative feedback controllers for robust perfect adaptation and dynamic performance. Nat Commun 13:2119
doi: 10.1038/s41467-022-29640-7
pubmed: 35440114
pmcid: 9018779
Schroda M, Blöcker D, Beck CF (2000) The HSP70A promoter as a tool for the improved expression of transgenes in Chlamydomonas. Plant J 21:121–131
doi: 10.1046/j.1365-313x.2000.00652.x
pubmed: 10743653
Scranton MA, Ostrand JT, Georgianna DR, Lofgren SM, Li D, Ellis RC, Carruthers DN, Dräger A, Masica DL, Mayfield SP (2016) Synthetic promoters capable of driving robust nuclear gene expression in the green alga Chlamydomonas reinhardtii. Algal Res 15:135–142
doi: 10.1016/j.algal.2016.02.011
McQuillan JL, Berndt AJ, Sproles AE, Mayfield SP, Pandhal J (2022) Novel cis-regulatory elements as synthetic promoters to drive recombinant protein expression from the Chlamydomonas reinhardtii nuclear genome. Nat Biotechnol 25:9–18
Crozet P, Navarro FJ, Willmund F, Mehrshahi P, Bakowski K, Lauersen KJ, Pérez-Pérez ME, Auroy P, Gorchs Rovira A, Sauret-Gueto S, Niemeyer J, Spaniol B, Theis J, Trösch R, Westrich LD, Vavitsas K, Baier T, Hübner W, de Carpentier F, Cassarini M, Danon A, Henri J, Marchand CH, de Mia M, Sarkissian K, Baulcombe DC, Peltier G, Crespo JL, Kruse O, Jensen PE, Schroda M, Smith AG, Lemaire SD (2018) Birth of a photosynthetic chassis: a MoClo toolkit enabling synthetic biology in the microalga Chlamydomonas reinhardtii. ACS Synth Biol 7:2074–2086
doi: 10.1021/acssynbio.8b00251
pubmed: 30165733
Weber E, Engler C, Gruetzner R, Werner S, Marillonnet S (2011) A modular cloning system for standardized assembly of multigene constructs. PLoS One 18(6):e16765
doi: 10.1371/journal.pone.0016765
Niemeyer J, Schroda M (2022) New destination vectors facilitate modular cloning for Chlamydomonas. Curr Genet 68:531–536
doi: 10.1007/s00294-022-01239-x
pubmed: 35429260
pmcid: 9279246