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Targeted delivery of fluorogenic peptide aptamers into live microalgae by femtosecond laser photoporation at single-cell resolution

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dc.contributor.author Maeno, Takanori en
dc.contributor.author Uzawa, Takanori en
dc.contributor.author Kono, Izumi en
dc.contributor.author Okano, Kazunori en
dc.contributor.author Iino, Takanori en
dc.contributor.author Fukita, Keisuke en
dc.contributor.author Oshikawa, Yuki en
dc.contributor.author Ogawa, Taro en
dc.contributor.author Iwata, Osamu en
dc.contributor.author Ito, Takuro en
dc.contributor.author Suzuki, Kengo en
dc.contributor.author Goda, Keisuke en
dc.contributor.author Hosokawa, Yoichiroh en
dc.date.accessioned 2018-06-21T06:29:13Z en
dc.date.available 2018-06-21T06:29:13Z en
dc.date.issued 2018-05-29 en
dc.identifier.issn 2045-2322 en
dc.identifier.uri http://hdl.handle.net/10061/12555 en
dc.description.abstract Microalgae-based metabolic engineering has been proven effective for producing valuable substances such as food supplements, pharmaceutical drugs, biodegradable plastics, and biofuels in the past decade. The ability to accurately visualize and quantify intracellular metabolites in live microalgae is essential for efficient metabolic engineering, but remains a major challenge due to the lack of characterization methods. Here we demonstrate it by synthesizing fluorogenic peptide aptamers with specific binding affinity to a target metabolite and delivering them into live microalgae by femtosecond laser photoporation at single-cell resolution. As a proof-of-principle demonstration of our method, we use it to characterize Euglena gracilis, a photosynthetic unicellular motile microalgal species, which is capable of producing paramylon (a carbohydrate granule similar to starch). Specifically, we synthesize a peptide aptamer containing a paramylon-binding fluorescent probe, 7-nitrobenzofurazan, and introduce it into E. gracilis cells one-by-one by suppressing their mobility with mannitol and transiently perforating them with femtosecond laser pulses at 800nm for photoporation. To demonstrate the method’s practical utility in metabolic engineering, we perform spatially and temporally resolved fluorescence microscopy of single live photoporated E. gracilis cells under different culture conditions. Our method holds great promise for highly efficient microalgae-based metabolic engineering. ja
dc.language.iso en en
dc.publisher Nature Publishing Group UK en
dc.rights © 2018 Macmillan Publishers Limited, part of Springer Nature ja
dc.title Targeted delivery of fluorogenic peptide aptamers into live microalgae by femtosecond laser photoporation at single-cell resolution en
dc.type.nii Journal Article en
dc.contributor.transcription ホソカワ, ヨウイチロウ ja
dc.contributor.alternative 細川, 陽一郎 ja
dc.textversion publisher en
dc.identifier.abstracturl https://www.nature.com/articles/s41598-018-26565-4#Abs1 en
dc.identifier.jtitle Scientific Reports en
dc.identifier.volume 8 en
dc.identifier.issue 1 en
dc.relation.doi 10.1038/s41598-018-26565-4 en
dc.identifier.artnum 8271 en
dc.identifier.NAIST-ID 73295875 en
dc.relation.pmid 29844463 en

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