The DOI matches its Crossref record; 19 of the 19 references it cites were found in a registry or an index.
A Modular Cloning System for Standardized Assembly of Multigene Constructs
Open accessAbstract
The field of synthetic biology promises to revolutionize biotechnology through the design of organisms with novel phenotypes useful for medicine, agriculture and industry. However, a limiting factor is the ability of current methods to assemble complex DNA molecules encoding multiple genetic elements in various predefined arrangements. We present here a hierarchical modular cloning system that allows the creation at will and with high efficiency of any eukaryotic multigene construct, starting from libraries of defined and validated basic modules containing regulatory and coding sequences. This system is based on the ability of type IIS restriction enzymes to assemble multiple DNA fragments in a defined linear order. We constructed a 33 kb DNA molecule containing 11 transcription units made from 44 individual basic modules in only three successive cloning steps. This modular cloning (MoClo) system can be readily automated and will be extremely useful for applications such as gene stacking and metabolic engineering.
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Cites
Enzymatic assembly of DNA molecules up to several hundred kilobases
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Self-citationSynthetic biology: applications come of age
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Golden Gate Shuffling: A One-Pot DNA Shuffling Method Based on Type IIs Restriction Enzymes
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Self-citationRefinement and standardization of synthetic biological parts and devices
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A CRISPR/Cas9 toolkit for multiplex genome editing in plants
Xing, H.-L., Dong, L., Wang, Z.-P., Zhang, H.-Y., Han, C.-Y., Liu, B., Wang, X.-C., & Chen, Q.-J. (2014) · BMC Plant Biology · doi:10.1186/s12870-014-0327-y · Cited by 1,684
A Highly Characterized Yeast Toolkit for Modular, Multipart Assembly
Lee, M. E., DeLoache, W. C., Cervantes, B., & Dueber, J. E. (2015) · ACS Synthetic Biology · doi:10.1021/sb500366v · Cited by 916
Large-scale de novo DNA synthesis: technologies and applications
Kosuri, S., & Church, G. M. (2014) · Nature Methods · doi:10.1038/nmeth.2918 · Cited by 869
Lawrenson, T., Shorinola, O., Stacey, N., Li, C., Østergaard, L., Patron, N., Uauy, C., & Harwood, W. (2015) · Genome Biology · doi:10.1186/s13059-015-0826-7 · Cited by 551
Munoz, D. M., Cassiani, P. J., Li, L., Billy, E., Korn, J. M., Jones, M. D., Golji, J., Ruddy, D. A., Yu, K., McAllister, G., et al. (2016) · Cancer Discovery · doi:10.1158/2159-8290.cd-16-0178 · Cited by 359
Metadata
- Authors
- Weber, E., Engler, C., Gruetzner, R., Werner, S., & Marillonnet, S.
- Venue
- PLoS ONE
- Publisher
- Public Library of Science (PLoS)
- Published
- 2011-02-18
- Volume
- 6
- Issue
- 2
- Pages
- e16765
- ISSN
- 1932-6203 (ISSN-L)
- OpenAlex
- W1970893992
- Cited by
- 1,377
- References
- 19
- FWCI
- 9.99
- Type
- journal article
- Language
- en
- Open access
- gold
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