Reproducibility of fluorescent expression from engineered biological constructs in E. coli

Jacob Beal, Traci Haddock-Angelli, Markus Gershater, Kim De Mora, Meagan Lizarazo, Jim Hollenhorst, Randy Rettberg, Philipp Demling, Rene Hanke, Michael Osthege, Anna Schechtel, Suresh Sudarsan, Arne Zimmermann, Bartosz Gabryelczyk, Martina Ikonen, Minnamari Salmela, Muradýye Acar, Muhammed Fatih Aktas, Furkan Bestepe, Furkan Sacit CeylanSadýk Cigdem, Mikail Dohan, Mustafa Elitok, Mehmet Gunduz, Esra Gunduz, Omer Faruk Hatipoglu, Turan Kaya, Orhan Sayin, Safa Tapan, Osman Faruk Tereci, Abdullah Uçar, Mustafa Yilmaz, Jeffrey Barrick, Alex Gutierrez, Dennis Mishler, Jordan Monk, Kate Mortensen, Nathan Shin, Ella Watkins, Yintong Chen, Yuji Jin, Yuanjie Shi, Haoqian Myelin Zhang, Bruno Ono, Ieda Maria Martinez Paino, Lais Ribovski, Ivan Silva, Danilo Keiji Zampronio, Nils Birkholz, Rudiger Frederik Busche, Oliver Konzock, Steffen Lippold, Carsten Ludwig, Melanie Philippi, Lukas Platz, Christian Sigismund, Susanne Weber, Maren Wehrs, Niels Werchau, Anna Wronska, Zen Zen Yen, Yash Agarwal, Evan Appleton, Douglas Densmore, Ariela Esmurria, Kathleen Lewis, Alan Pacheco, Marcel Bruchez, Danielle Peters, Cheryl Telmer, Lena Wang, Silvia Canas-Duarte, Daniel Giraldo-Perez, Camilo Gomez-Garzon, Jorge Madrid-Wolff, Nathaly Marin-Medina, Valentina Mazzanti, Laura Rodriguez-Forero, Eitan Scher, Robin Dowell, Samantha O'Hara, Cloe Simone Pogoda, Kendra Shattuck, Ali Altintas, Anne Pihl Bali, Rasmus Bech, Anne Egholm, Anne Sofie Laerke Hansen, Kristian Jensen, Kristian Barreth Karlsen, Caroline Mosbech, Sophia Belkhelfa, Noemie Berenger, Romain Bodinier, Cecile Jacry, Laura Matabishi-Bibi, Pierre Parutto, Julie Zaworski, Andries De Vries, Freek De Wijs, Rick Elbert, Lisa Hielkema, Chandhuru Jagadeesan, Bayu Jayawardhana, Oscar Kuipers, Anna Lauxen, Thomas Meijer, Sandra Mous, Renske Van Raaphorst, Aakanksha Saraf, Otto Schepers, Oscar Smits, Jan Willem Veening, Ruud Detert Oude Weme, Lianne Wieske, Catherine Ainsworth, Xenia Spencer-Milnes, Alejandro Gómez Ávila, Eddie Cano Gamez, Ana Laura Torres Huerta, Carlos Alejandro Meza Ramirez, Philipp Popp, Jara Radeck, Anna Sommer, Xiangkai Li, Qi Wu, Hongxia Zhao, Ruixue Zhao, Irem Bastuzel, Yasemin Ceyhan, Mayda Gursel, Burak Kizil, Ilkem Kumru, Yasemin Kuvvet, Helin Tercan, Seniz Yuksel, Luiza Niyazmetova, Timothy Ang, Lucas Black, Ciaran Kelly, George Wadhams, Clovis Basier, Urszula Czerwinska, Cindy Suci Ananda, Muhammad Al Azhar, Adelia Elviantari, Maya Fitriana, Arief Budi Witarto, Yulianti, Qingfeng Hou, Wan Pei, Chen Rifei, Wang Rong, Huang Wei, Zhang Yushan, He Jianguo, Dengwen Lai, Pai Li, Jianheng Liu, Chunyang Ni, Qianbin Zhang, Cinthya Cadenas, Eduardo J Zardain Canabal, Claudia Nallely Alonso Cantu, Mercedes Alejandra Vazquez Cantu, Eduardo Cepeda Canedo, Cesar Miguel Valdez Cordova, Jose Alberto De La Paz Espinosa, Carlos Enrique Alavez Garcia, Ana Laura Navarro Heredia, Adriana Hernandez, Sebastian Valdivieso Jauregui, Eduardo Ramirez Montiel, Eduardo Serna Morales, Yamile Minerva Castellanos Morales, Omar Alonso Cantu Pena, Eduardo A. Ramirez-Rodríguez, Elizabeth Vallejo Trejo, Jesus Gilberto Rodriguez Ceja, Jesus Eduardo Martinez Hernandez, Mario Alberto Pena Hernandez, Enrique Amaya Perez, Rebeca Paola Torres Ramirez, J. Claudio Moreno Rocha, Alber Sanchez, Claudia Melissa Guerra Vazquez, Martin Hanzel, Sarah Mohand-Said, Shihab Sawar, Dylan Siriwardena, Alex Tzahristos, Nils Anlind, Martin Friberg, Erik Gullberg, Stephanie Herman, Dallin Christensen, Sara Gertsch, Cody Maxfield, Charles Miller, Ryan Putman, Christine Bauerl, Lucia T Estelles Lopez, Estefania Huet-Trujillo, Marta Vazquez Vilar, Marlène Sophie Birk, Nico Claassens, Walter De Koster, Rik Van Rosmalen, Wen Ying Wu, Sian Davies, Dan Goss, William Rostain, Chelsey Tye, Waqar Yousaf, Natalie Farny, Chloe LaJeunesse, Alex Turland, An Chen, Jielin Chen, Yahong Chen, Zehua Chen, Baishan Fang, Xiaotong Fu, Xifeng Guo, Yiying Lei, Jianqiao Li, Zhe Li, Chang Liu, Weibing Liu, Yang Liu, Yizhu Lv, Qingyu Ruan, Yue Su, Chun Tang, Yushen Wang, Fan Wu, Xiaoshan Yan, Ruihua Zhang, Tangduo Zhang, Farren Isaacs, Ariel Leyva-Hernandez, Natalie Ma, Stephanie Mao, Yamini Naidu, Tuukka Miinalainen, Marion Aruanno, Daniel Calendini, Yoann Chabert, Gael Chambonnier, Myriam Choukour, Ella De Gaulejac, Camille Houy, Axel Levier, Loreen Logger, Sebastien Nin, Valerie Prima, James N. Sturgis, Beibei Fang, Sadik Cigdem, Alejandro Gutierrez, Revanth Poondla, Sanjana Reddy, Tyler Rocha, Natalie Schulte, Devin Wehle, Marta Eva Jackowski, Sean Ross Craig, Ariana Mirzarafie-Ahi, Elliott Parris, Luba Prout, Barbara Steijl, Rachel Wellman, Zhao Fan, Zhang Jing, Yang Wei, Yang Yuanzhan, Wen Zhaosen, Evan Appletion, Jeffrey Chen, Abha Patil, Shaheer Priracha, Kate Ryan, Nick Salvador, John Viola, Camila Maria S Boralli, Camila Barbosa Bramorski, Juliana Cancino-Bernardi, Ana Laura De Lima, Paula Maria Pincela Lins, Cristiane Casonato Melo, Deborah Cezar Mendonca, Thiago Mosqueiro, Everton Silva, Graziele Vasconcelos, Ruchi Asthana, Donna Lee, Michelle Yu, Peter Choi, Effie Lau, Kenneth Lau, Oscar Ying, Brandon Malone, Paul Young, Aidan Ceney, Dakota Hawthorne, Sharon Lian, Sam Mellentine, Dylan Miller, Barbara Castro Moreira, Christie Peebles, Olivia Smith, Kevin Walsh, Allison Zimont, Michael Brasino, Michael Donovan, Hannah Young, Jan Bejvl, Daniel Georgiev, Hynek Kasl, Katerina Pechotova, Vaclav Pelisek, Anna Sosnova, Pavel Zach, Anthony Ciesla, Benjamin Hoover, Elliott Chapman, Jon Marles-Wright, Vicky Moynihan, Liusaidh Owen, Brooke Rothschild-Mancinelli, Emilie Cuillery, Joseph Heng, Vincent Jacquot, Paola Malsot, Rocco Meli, Cyril Pulver, Ari Sarfatis, Loic Steiner, Victor Steininger, Nina Van Tiel, Gregoire Thouvenin, Axel Uran, Lisa Baumgartner, Anna Fomitcheva, Daniel Gerngross, Verena Jagger, Michael Meier, Anja Michel, Jasmine Bird, Bradley Brown, Todd Burlington, Daniel Herring, Joseph Slack, Georgina Westwood, Emilia Wojcik, Julian Bender, Julia Donauer, Ramona Emig, Rabea Jesser, Julika Neumann, Lara Stuhn, Takema Hasegawa, Tomoya Kozakai, Haruka Maruyama, Sean Colloms, Charlotte Flynn, Vilija Lomeikaite, James Provan, Kang Ning, Shuyan Tang, Guozhao Wu, Yunjun Yang, Zhi Zeng, Yi Zhan, Pan Chu, Jun Li, Keji Yan, Chaitanya A. Athale, Swapnil Bodkhe, Manasi Gangan, Harsh Gakhare, Yash Jawale, Snehal Kadam, Prachiti Moghe, Gayatri Mundhe, Neha Khetan, Ira Phadke, Prashant Uniyal, Siddhesh Zadey, Ines Cottignie, Eline Deprez, Astrid Deryckere, Jasper Janssens, Frederik Jonnaert, Katarzyna Malczewska, Thomas Pak, Johan Robben, Ovia Margaret Thirukkumaran, Vincent Van Deuren, Laurens Vandebroek, Laura Van Hese, Laetitia Van Wonterghem, Leen Verschooten, Moritz Wolter, Joss Auty, Richard Badge, Liam Crawford, Raymond Dalgleish, Amy Evans, Cameron Grundy, Charlie Kruczko, Payal Karia, Graeme Glaister, Rhys Hakstol, Seme Mate, Karin Otero, Dustin Smith, Jeff Tingley, Hans Joachim Wieden, Haotian Wang, Ningning Yao, Matthias Franz, Anna Knoerlein, Nicolas Koutsoubelis, Anne C. Loechner, Max Mundt, Alexandra Richter, Oliver Schauer, Marjorie Buss, Sivateja Tangirala, Brian Teague, Tianyi Huang, Xinhao Song, Yibing Wei, Zhaoran Zhang, Longzhi Cao, Cheng Li, Kang Yang, Zhiqin Chen, Yuxing Fang, Libo Sun, Weiyi Wang, Yang Yang, David Adams, Joshua Colls, Joshua Timmons, David Urick, Julia Anna Adrian, Madina Akan, Youssef Chahibi, Rahmi Lale, Typhaine Le Doujet, Marit Vaagen Roee, Altynay Abdirakhmanova, Askarbek Orakov, Azhar Zhailauova, Jinyang Liang, Yu Ma, Qikai Qin, Yetian Su, Ju Yeon Han, Raphaella Hull, Wei Chung Kong, Li Chieh Lu, Duke Quinton, Pauline Aubert, Johan Bourdarias, Olivier Bugaud, Coralie Demon-Chaine, Isabelle Hatin, Ibtissam Kaid-Slimane, Seong Koo Kang, Audrey Moatti, Cheikh Fall Ndiaye, Mathilde Ananos, Alexander Arkhipenko, Valentin Bailly, Jules Caput, Javier Castillo, Alma Chapet-Batlle, Floriane Cherrier, Claudia Demarta-Gatsi, Deshmukh Gopaul, Muriel Gugger, Caroline Lambert, Lucas Krauss, Amelie Vandendaele, Li Xiaojing, Lin Xiaomei, Luo Xunxun, Anders Chr Hansen, Tina Kronborg, Jens S. Pettersen, Charles Calvet, Tyler Dae Devlin, Kosuke Fujishima, Danny Greenberg, Tina Ju, Ryan Kent, Daniel Kunin, Erica Lieberman, Griffin McCutcheon, Thai Nguyen, Lynn Rothschild, Joseph D. Shih, Jack Takahashi, Kirsten Thompson, Forrest Tran, Daniel Xiang, Felix Richter, Yang Xiaoran, Hu Xiangyue, Changyuan Deng, Shuyu Hua, Yumeng Li, Xinyu Meng, Boxiang Wang, Yingqi Wang, Xuan Wang, Zixuan Xu, Jieyu Yan, Ming Yan, Yineng Zhou, Edgar Alberto Alcalá Orozco, José Alberto Cristerna Bermúdez, Daniela Flores Gómez, José Ernesto Hernández Castañeda, Diana Clarisse Montaño Navarro, Juana Yessica Pérezávila, María Fernanda Salazar Figueroa, María Fernanda Sánchez Arroyo, Oliva Angélica Sánchez Montesinos, Ángel Farid Rojas Cruz, Carlos Ramos Gutiérrez, Alonso Pérez Lona, Fernanda Sotomayor Olivares, Jorge Sebastián Rodríguez Iniesta, Juan Carlos Rueda Silva, Shotaro Ayukawa, Takahiro Kashiwagi, Daisuke Kiga, Misa Minegishi, Riku Shinohara, Hiraku Tokuma, Yuta Yamazaki, Shuhei Yasunishi, Erinn Sim Zixuan, Remsha Afzal, Matthew Carrigan, Barry Moran, Marlena Mucha, Arnas Petrauskas, Stefan Marsden, Michelle Post, Anne Rodenburg, Hector Sanguesa, Marit Van Der Does, Erwin Van Rijn, Max Van't Hof, Yeshi De Bruin, Hans De Ferrante, Elles Elschot, Laura Jacobs, Jan Willem Muller, Sjoerd Nooijens, Femke Vaassen, Cas Van Der Putten, Esther Van Leeuwen, Laura Van Smeden, Kwan Kwan Zhu, Kevin Sabath, Katharina Sporbeck, Nicolai Von Kügelgen, Lisa Wellinger, Stefanie Braun, Jack Ho, Yash Mishra, Mariola Sebastian, Lucas Von Chamier, Fasih M. Ahsan, Megan A. Satyadi, Vivienne Gunadhi, Phillip Kyriakakis, Jenny Lee, Walter Thavarajah, Kimia Abtahi, Robert Hand, Chun Mun Loke, Adam Wahab, Iowis Zhu, Cristina Del Bianco, Fabio Chizzolini, Elisa Godino, Roberta Lentini, Sheref S. Mansy, Noel Yeh Martin, Claudio Oss Pegorar, Alexander Cook, Timothy Kerns, Chad Nielsen, Michael Paskett, Alexander Torgesen, Stephen Lee, Ophir Ospovat, Sikandar Raza, Daniel Shaykevich, Jarrod Shilts, Barbora Bajorinaite, Mykolas Bendorius, Ieva Rauluseviciute, Ieva Savickyte, Sarunas Tumas, William Buchser, Elli Cryan, Caroline Golino, Andrew Halleran, Taylor Jacobs, Michael LeFew, Joe Maniaci, John Marken, Margaret Saha, Panya Vij, Kayla DeSanty, Julie Mazza, Yue Jiang, Yang Liu, Yang Yang

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Abstract

We present results of the first large-scale interlaboratory study carried out in synthetic biology, as part of the 2014 and 2015 International Genetically Engineered Machine (iGEM) competitions. Participants at 88 institutions around the world measured fluorescence from three engineered constitutive constructs in E. coli. Few participants were able to measure absolute fluorescence, so data was analyzed in terms of ratios. Precision was strongly related to fluorescent strength, ranging from 1.54-fold standard deviation for the ratio between strong promoters to 5.75-fold for the ratio between the strongest and weakest promoter, and while host strain did not affect expression ratios, choice of instrument did. This result shows that high quantitative precision and reproducibility of results is possible, while at the same time indicating areas needing improved laboratory practices.

Original languageEnglish
Article numbere0150182
Pages (from-to)1-22
JournalPloS one
Volume11
Issue number3
DOIs
Publication statusPublished - 1 Mar 2016
MoE publication typeA1 Journal article-refereed

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