Team:ZJU-China/Introduction

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Why circuit construction?

Genetic regulatory circuits are artificially-designed gene clusters constructed by disparate genetic elements, which can produce novel genetic function according to people’s desire. Able to be modeled and simulated in silico, genetic regulatory circuits can be either qualitatively or quantitatively examined, with its function even able to be predicted. Sound construction of circuits has been one of the crucial parts of synthetic biology, which has been taking people great efforts to perfect.

Why on chromosome?

Nowadays, modern techniques have enabled the construction of more complicated and large-capacity genetic systems. However, most of the genetic circuits are constructed on plasmids, which, with the increasing of complexity, has brought about incremental uncertainty and unpredictability. This indeterminacy is mainly generated by plasmid loss, allele inactivation, copy number variability or plasmid-associated metabolic burden[1]. To obtain optimal performance in certain microbial host, rounds of examination and troubleshooting may be needed. Nevertheless, with genetic regulatory circuits constructed on microbial chromosome or on bacterial artificial chromosome (BAC), more robust systems can be obtained.

Why GeneSocket?

With more and more complicated genetic circuits designed, more efficient, time-saving and inexpensive methods are needed to bring the design into reality. Many well-known methods like traditional restriction digestion and ligation, 3A assembly and Gibson assembly all aim to overcome the difficulties of gene assembly while problems like cumbersome steps, low cost performance, lots of time consuming do not receive quite effective solutions. The new gene-insertion method we build, which we call GeneSocket, is able to assembly genetic elements in vivo efficiently, with reporters easy to be identified, and simple isolation methods. We hope that it can become another choice for researchers in future.

Reference

[1]Santos, C. N. S. & Yoshikuni, Y. Engineering complex biological systems in bacteria through recombinase-assisted genome engineering. Nature Protocols 9, 1320-1336, doi:10.1038/nprot.2014.084 (2014).