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Contents Vol 63(9)

Microfluidic Droplet Technique for In Vitro Directed Evolution

Nan Wu A B , John Oakeshott C , Sue Brown D , Christopher Easton B E and Yonggang Zhu A E
+ Author Affiliations
- Author Affiliations

A CSIRO Materials Science and Engineering, PO Box 56, Highett, Vic. 3190, Australia.

B Research School of Chemistry, Australian National University, ACT 0200, Australia.

C CSIRO Entomology, GPO Box 1700, Acton, Canberra, ACT 2601, Australia.

D CSIRO Corporate Centre, PO Box 225, Dickson, ACT 2602, Australia.

E Corresponding authors. Email: yonggang.zhu@csiro.au; easton@rsc.anu.edu.au

Australian Journal of Chemistry 63(9) 1313-1325 https://doi.org/10.1071/CH10116
Submitted: 10 March 2010  Accepted: 8 June 2010   Published: 9 September 2010

Abstract

Increasingly over the past two decades, biotechnologists have been exploiting various molecular technologies for high-throughput screening of genes and their protein products to isolate novel functionalities with a wide range of industrial applications. One particular technology now widely used for these purposes involves directed evolution, an artificial form of evolution in which genes and proteins are evolved towards new or improved functions by imposing intense selection pressures on libraries of mutant genes generated by molecular biology techniques and expressed in heterologous systems such as Escherichia coli. Most recently, the rapid development of droplet-based microfluidics has created the potential to dramatically increase the power of directed evolution by increasing the size of the libraries and the throughput of the screening by several orders of magnitude. Here, we review the methods for generating and controlling droplets in microfluidic systems, and their applications in directed evolution. We focus on the methodologies for cell-based assays, in vitro protein expression and DNA amplification, and the prospects for using such platforms for directed evolution in next-generation biotechnologies.


Acknowledgements

N.W. gratefully acknowledges financial support from The Australian National University through the post-graduate scholarship scheme and CSIRO’s Synthetic Enzymes Emerging Science Initiative.


References


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