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Title: Droplet microfluidics for synthetic biology

Here, synthetic biology is an interdisciplinary field that aims to engineer biological systems for useful purposes. Organism engineering often requires the optimization of individual genes and/or entire biological pathways (consisting of multiple genes). Advances in DNA sequencing and synthesis have recently begun to enable the possibility of evaluating thousands of gene variants and hundreds of thousands of gene combinations. However, such large-scale optimization experiments remain cost-prohibitive to researchers following traditional molecular biology practices, which are frequently labor-intensive and suffer from poor reproducibility. Liquid handling robotics may reduce labor and improve reproducibility, but are themselves expensive and thus inaccessible to most researchers. Microfluidic platforms offer a lower entry price point alternative to robotics, and maintain high throughput and reproducibility while further reducing operating costs through diminished reagent volume requirements. Droplet microfluidics have shown exceptional promise for synthetic biology experiments, including DNA assembly, transformation/transfection, culturing, cell sorting, phenotypic assays, artificial cells and genetic circuits.
Authors:
 [1] ;  [1] ;  [1] ;  [2] ; ORCiD logo [1]
  1. DOE Joint BioEnergy Institute, Emeryville, CA (United States); Sandia National Lab. (SNL-CA), Livermore, CA (United States)
  2. DOE Joint BioEnergy Institute, Emeryville, CA (United States); Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States); USDOE Joint Genome Institute (JGI), Walnut Creek, CA (United States)
Publication Date:
Report Number(s):
SAND-2017-9052J
Journal ID: ISSN 1473-0197; LCAHAM; 656492
Grant/Contract Number:
AC04-94AL85000
Type:
Accepted Manuscript
Journal Name:
Lab on a chip (Print)
Additional Journal Information:
Journal Name: Lab on a chip (Print); Journal Volume: 17; Journal Issue: 20; Journal ID: ISSN 1473-0197
Publisher:
Royal Society of Chemistry
Research Org:
Sandia National Lab. (SNL-NM), Albuquerque, NM (United States)
Sponsoring Org:
USDOE Office of Science (SC), Biological and Environmental Research (BER) (SC-23)
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES
OSTI Identifier:
1421856

Gach, Philip Charles, Iwai, Kosuke, Kim, Peter Wonhee, Hillson, Nathan J., and Singh, Anup K.. Droplet microfluidics for synthetic biology. United States: N. p., Web. doi:10.1039/C7LC00576H.
Gach, Philip Charles, Iwai, Kosuke, Kim, Peter Wonhee, Hillson, Nathan J., & Singh, Anup K.. Droplet microfluidics for synthetic biology. United States. doi:10.1039/C7LC00576H.
Gach, Philip Charles, Iwai, Kosuke, Kim, Peter Wonhee, Hillson, Nathan J., and Singh, Anup K.. 2017. "Droplet microfluidics for synthetic biology". United States. doi:10.1039/C7LC00576H. https://www.osti.gov/servlets/purl/1421856.
@article{osti_1421856,
title = {Droplet microfluidics for synthetic biology},
author = {Gach, Philip Charles and Iwai, Kosuke and Kim, Peter Wonhee and Hillson, Nathan J. and Singh, Anup K.},
abstractNote = {Here, synthetic biology is an interdisciplinary field that aims to engineer biological systems for useful purposes. Organism engineering often requires the optimization of individual genes and/or entire biological pathways (consisting of multiple genes). Advances in DNA sequencing and synthesis have recently begun to enable the possibility of evaluating thousands of gene variants and hundreds of thousands of gene combinations. However, such large-scale optimization experiments remain cost-prohibitive to researchers following traditional molecular biology practices, which are frequently labor-intensive and suffer from poor reproducibility. Liquid handling robotics may reduce labor and improve reproducibility, but are themselves expensive and thus inaccessible to most researchers. Microfluidic platforms offer a lower entry price point alternative to robotics, and maintain high throughput and reproducibility while further reducing operating costs through diminished reagent volume requirements. Droplet microfluidics have shown exceptional promise for synthetic biology experiments, including DNA assembly, transformation/transfection, culturing, cell sorting, phenotypic assays, artificial cells and genetic circuits.},
doi = {10.1039/C7LC00576H},
journal = {Lab on a chip (Print)},
number = 20,
volume = 17,
place = {United States},
year = {2017},
month = {8}
}

Works referenced in this record:

Enzymatic assembly of DNA molecules up to several hundred kilobases
journal, April 2009
  • Gibson, Daniel G.; Young, Lei; Chuang, Ray-Yuan
  • Nature Methods, Vol. 6, Issue 5, p. 343-345
  • DOI: 10.1038/nmeth.1318

Deoxynucleoside phosphoramidites—A new class of key intermediates for deoxypolynucleotide synthesis
journal, January 1981

j5 DNA Assembly Design Automation Software
journal, December 2011
  • Hillson, Nathan J.; Rosengarten, Rafael D.; Keasling, Jay D.
  • ACS Synthetic Biology, Vol. 1, Issue 1, p. 14-21
  • DOI: 10.1021/sb2000116

Ultrahigh-throughput screening in drop-based microfluidics for directed evolution
journal, February 2010
  • Agresti, J. J.; Antipov, E.; Abate, A. R.
  • Proceedings of the National Academy of Sciences, Vol. 107, Issue 9, p. 4004-4009
  • DOI: 10.1073/pnas.0910781107

Scalable gene synthesis by selective amplification of DNA pools from high-fidelity microchips
journal, November 2010
  • Kosuri, Sriram; Eroshenko, Nikolai; LeProust, Emily M.
  • Nature Biotechnology, Vol. 28, Issue 12, p. 1295-1299
  • DOI: 10.1038/nbt.1716

A One Pot, One Step, Precision Cloning Method with High Throughput Capability
journal, November 2008