Protein Synthesis via in vitro Transcription and Translation System inside Monodisperse Vesicles Fabricated by Microfluidics
ABSTRACT In this study, we investigated the conditions for protein synthesis by an in vitro transcription and translation (IVTT) system within giant unilamellar vesicles (GUVs) produced with a microfluidic channel. The commercial IVTT system consisted of purified components (PURE system), and DNA en...
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| Published in | Electrical engineering in Japan Vol. 218; no. 2 |
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| Main Authors | , , , |
| Format | Journal Article |
| Language | English |
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| Abstract | ABSTRACT
In this study, we investigated the conditions for protein synthesis by an in vitro transcription and translation (IVTT) system within giant unilamellar vesicles (GUVs) produced with a microfluidic channel. The commercial IVTT system consisted of purified components (PURE system), and DNA encoding target protein was encapsulated in GUVs and incubated to synthesize the proteins. Synthesis of green fluorescent protein (GFP) and nanopore‐forming α‐hemolysin were tested as the models of water‐soluble protein and membrane protein, respectively. The stability of the GUVs and the efficiency of protein synthesis were assessed, focusing on variations in the concentration of the PURE system and the size of the GUVs. Our findings contribute to the development of homogeneous bioreactors and biosensors based on GUV technology. |
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| AbstractList | In this study, we investigated the conditions for protein synthesis by an in vitro transcription and translation (IVTT) system within giant unilamellar vesicles (GUVs) produced with a microfluidic channel. The commercial IVTT system consisted of purified components (PURE system), and DNA encoding target protein was encapsulated in GUVs and incubated to synthesize the proteins. Synthesis of green fluorescent protein (GFP) and nanopore‐forming α‐hemolysin were tested as the models of water‐soluble protein and membrane protein, respectively. The stability of the GUVs and the efficiency of protein synthesis were assessed, focusing on variations in the concentration of the PURE system and the size of the GUVs. Our findings contribute to the development of homogeneous bioreactors and biosensors based on GUV technology. ABSTRACT In this study, we investigated the conditions for protein synthesis by an in vitro transcription and translation (IVTT) system within giant unilamellar vesicles (GUVs) produced with a microfluidic channel. The commercial IVTT system consisted of purified components (PURE system), and DNA encoding target protein was encapsulated in GUVs and incubated to synthesize the proteins. Synthesis of green fluorescent protein (GFP) and nanopore‐forming α‐hemolysin were tested as the models of water‐soluble protein and membrane protein, respectively. The stability of the GUVs and the efficiency of protein synthesis were assessed, focusing on variations in the concentration of the PURE system and the size of the GUVs. Our findings contribute to the development of homogeneous bioreactors and biosensors based on GUV technology. |
| Author | Suzuki, Hiroaki Tsugane, Mamiko Nanjo, Satoshi Matsuura, Tomoaki |
| Author_xml | – sequence: 1 givenname: Satoshi surname: Nanjo fullname: Nanjo, Satoshi organization: Chuo University – sequence: 2 givenname: Mamiko surname: Tsugane fullname: Tsugane, Mamiko organization: Chuo University – sequence: 3 givenname: Tomoaki surname: Matsuura fullname: Matsuura, Tomoaki organization: Institute of Future Science, Institute of Science Tokyo – sequence: 4 givenname: Hiroaki surname: Suzuki fullname: Suzuki, Hiroaki email: suzuki@mech.chuo-u.ac.jp organization: Chuo University |
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| Cites_doi | 10.1021/acschembio.5b00107 10.1038/90802 10.1021/sb500249g 10.1021/jacs.6b02107 10.1021/acssynbio.3c00629 10.1021/sb400094c 10.1016/j.snb.2021.131281 10.1038/s41598-018-27547-2 10.1038/ncomms10447 10.1039/C1CS15211D |
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| Notes | Funding This study was supported by JSPS Grant‐in‐Aid for Scientific Research (19H02576, 19H00901, 20H05935, 21H05890, 21K04851, 22K21344, and 21H05228), Institute of Science and Engineering, Chuo University, and Human Frontier Science Program RGP003/2023. of IEEJ Transactions on Sensors and Micromachines 10.1541/ieejsmas.145.41 Translated from Volume 145 Number 3, pages 41–45, DOI (Denki Gakkai Ronbunshi E). ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 14 |
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| References_xml | – volume: 7 year: 2016 article-title: Octanol‐Assisted Liposome Assembly on Chip publication-title: Nature Communications – volume: 19 start-page: 751 issue: 8 year: 2001 end-page: 755 article-title: Cell‐Free Translation Reconstituted With Purified Components publication-title: Nature Biotechnology – volume: 355 year: 2022 article-title: Plug‐and‐Play Microfluidic Production of Monodisperse Giant Unilamellar Vesicles Using Droplet Transfer Across Water‐Oil Interface publication-title: Sensors and Actuators B: Chemical – volume: 3 start-page: 372 issue: 6 year: 2014 end-page: 379 article-title: In Vitro Membrane Protein Synthesis inside Cell‐Sized Vesicles Reveals the Dependence of Membrane Protein Integration on Vesicle Volume publication-title: ACS Synthetic Biology – volume: 10 start-page: 1694 issue: 7 year: 2015 end-page: 1701 article-title: Membrane Curvature Affects the Formation of α‐Hemolysin Nanopores publication-title: ACS Chemical Biology – volume: 13 start-page: 68 issue: 1 year: 2024 end-page: 76 article-title: Identifying Conditions for Protein Synthesis inside Giant Vesicles Using Microfluidics Toward Standardized Artificial Cell Production publication-title: ACS Synthetic Biology – volume: 8 start-page: 9214 year: 2018 article-title: Reverse Transcription Polymerase Chain Reaction in Giant Unilamellar Vesicles publication-title: Scientific Reports – volume: 138 start-page: 7584 issue: 24 year: 2016 end-page: 7591 article-title: Monodisperse Uni‐ and Multicompartment Liposomes publication-title: Journal of the American Chemical Society – volume: 4 start-page: 566 issue: 5 year: 2015 end-page: 576 article-title: Stochasticity in Gene Expression in a Cell‐Sized Compartment publication-title: ACS Synthetic Biology – volume: 41 start-page: 79 year: 2012 end-page: 85 article-title: Designs for Life: Protocell Models in the Laboratory publication-title: Chemical Society Reviews – ident: e_1_2_6_5_1 doi: 10.1021/acschembio.5b00107 – ident: e_1_2_6_10_1 doi: 10.1038/90802 – ident: e_1_2_6_4_1 doi: 10.1021/sb500249g – ident: e_1_2_6_7_1 doi: 10.1021/jacs.6b02107 – ident: e_1_2_6_11_1 doi: 10.1021/acssynbio.3c00629 – ident: e_1_2_6_6_1 doi: 10.1021/sb400094c – ident: e_1_2_6_9_1 doi: 10.1016/j.snb.2021.131281 – ident: e_1_2_6_3_1 doi: 10.1038/s41598-018-27547-2 – ident: e_1_2_6_8_1 doi: 10.1038/ncomms10447 – ident: e_1_2_6_2_1 doi: 10.1039/C1CS15211D |
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In this study, we investigated the conditions for protein synthesis by an in vitro transcription and translation (IVTT) system within giant... In this study, we investigated the conditions for protein synthesis by an in vitro transcription and translation (IVTT) system within giant unilamellar... |
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| SubjectTerms | Bioreactors Biosensors Fluorescence GUVs IVTT system liposome membrane protein Microfluidics protein Protein synthesis Proteins Vesicles |
| Title | Protein Synthesis via in vitro Transcription and Translation System inside Monodisperse Vesicles Fabricated by Microfluidics |
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