Imperial College London
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DrNaomiNakayama

Faculty of Engineering, Department of Bioengineering

Reader in Biological Design
 
 
 
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Contact

 

n.nakayama Website

 
 
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Location

 

B303ABessemer BuildingSouth Kensington Campus

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Summary

 

Publications

Citation

BibTex format

@article{Andreou:2018:10.1371/journal.pone.0189892,
author = {Andreou, AI and Nakayama, N},
doi = {10.1371/journal.pone.0189892},
journal = {PLoS One},
pages = {1--18},
title = {Mobius Assembly: A versatile Golden-Gate framework towards universal DNA assembly},
url = {http://dx.doi.org/10.1371/journal.pone.0189892},
volume = {13},
year = {2018}
}
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RIS format (EndNote, RefMan)

TY  - JOUR
AB  - Synthetic biology builds upon the foundation of engineering principles, prompting innovation and improvement in biotechnology via a design-build-test-learn cycle. A community-wide standard in DNA assembly would enable bio-molecular engineering at the levels of predictivity and universality in design and construction that are comparable to other engineering fields. Golden Gate Assembly technology, with its robust capability to unidirectionally assemble numerous DNA fragments in a one-tube reaction, has the potential to deliver a universal standard framework for DNA assembly. While current Golden Gate Assembly frameworks (e.g. MoClo and Golden Braid) render either high cloning capacity or vector toolkit simplicity, the technology can be made more versatile—simple, streamlined, and cost/labor-efficient, without compromising capacity. Here we report the development of a new Golden Gate Assembly framework named Mobius Assembly, which combines vector toolkit simplicity with high cloning capacity. It is based on a two-level, hierarchical approach and utilizes a low-frequency cutter to reduce domestication requirements. Mobius Assembly embraces the standard overhang designs designated by MoClo, Golden Braid, and Phytobricks and is largely compatible with already available Golden Gate part libraries. In addition, dropout cassettes encoding chromogenic proteins were implemented for cost-free visible cloning screening that color-code different cloning levels. As proofs of concept, we have successfully assembled up to 16 transcriptional units of various pigmentation genes in both operon and multigene arrangements. Taken together, Mobius Assembly delivers enhanced versatility and efficiency in DNA assembly, facilitating improved standardization and automation.
AU  - Andreou,AI
AU  - Nakayama,N
DO  - 10.1371/journal.pone.0189892
EP  - 18
PY  - 2018///
SN  - 1932-6203
SP  - 1
TI  - Mobius Assembly: A versatile Golden-Gate framework towards universal DNA assembly
T2  - PLoS One
UR  - http://dx.doi.org/10.1371/journal.pone.0189892
UR  - https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0189892
UR  - http://hdl.handle.net/10044/1/80966
VL  - 13
ER  -
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