An innovative platform for quick and flexible joining of assorted DNA fragments
Abstract
Successful synthetic biology efforts rely on conceptual and experimental designs in combination with testing of multi-gene constructs. Despite recent progresses, several limitations still hinder the ability to flexibly assemble and collectively share different types of DNA segments. We describe an advanced system for joining DNA fragments from a universal library that automatically maintains open reading frames (ORFs) and does not require linkers, adaptors, sequence homology, amplification or mutation (domestication) of fragments in order to work properly. Moreover, we find that this system, which is enhanced by a unique buffer formulation, provides unforeseen capabilities for testing, and sharing, complex multi-gene circuitry assembled from different DNA fragments.
- Authors:
-
- Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States). Biosciences Division
- Publication Date:
- Research Org.:
- Oak Ridge National Laboratory (ORNL), Oak Ridge, TN (United States)
- Sponsoring Org.:
- USDOE Office of Science (SC)
- OSTI Identifier:
- 1324162
- Alternate Identifier(s):
- OSTI ID: 1259839
- Grant/Contract Number:
- AC05-00OR22725; SC0008834
- Resource Type:
- Accepted Manuscript
- Journal Name:
- Scientific Reports
- Additional Journal Information:
- Journal Volume: 6; Journal ID: ISSN 2045-2322
- Publisher:
- Nature Publishing Group
- Country of Publication:
- United States
- Language:
- English
- Subject:
- 59 BASIC BIOLOGICAL SCIENCES
Citation Formats
De Paoli, Henrique Cestari, Tuskan, Gerald A., and Yang, Xiaohan. An innovative platform for quick and flexible joining of assorted DNA fragments. United States: N. p., 2016.
Web. doi:10.1038/srep19278.
De Paoli, Henrique Cestari, Tuskan, Gerald A., & Yang, Xiaohan. An innovative platform for quick and flexible joining of assorted DNA fragments. United States. https://doi.org/10.1038/srep19278
De Paoli, Henrique Cestari, Tuskan, Gerald A., and Yang, Xiaohan. Wed .
"An innovative platform for quick and flexible joining of assorted DNA fragments". United States. https://doi.org/10.1038/srep19278. https://www.osti.gov/servlets/purl/1324162.
@article{osti_1324162,
title = {An innovative platform for quick and flexible joining of assorted DNA fragments},
author = {De Paoli, Henrique Cestari and Tuskan, Gerald A. and Yang, Xiaohan},
abstractNote = {Successful synthetic biology efforts rely on conceptual and experimental designs in combination with testing of multi-gene constructs. Despite recent progresses, several limitations still hinder the ability to flexibly assemble and collectively share different types of DNA segments. We describe an advanced system for joining DNA fragments from a universal library that automatically maintains open reading frames (ORFs) and does not require linkers, adaptors, sequence homology, amplification or mutation (domestication) of fragments in order to work properly. Moreover, we find that this system, which is enhanced by a unique buffer formulation, provides unforeseen capabilities for testing, and sharing, complex multi-gene circuitry assembled from different DNA fragments.},
doi = {10.1038/srep19278},
journal = {Scientific Reports},
number = ,
volume = 6,
place = {United States},
year = {Wed Jan 13 00:00:00 EST 2016},
month = {Wed Jan 13 00:00:00 EST 2016}
}
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Cited by: 16 works
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Figures / Tables:
Figure 1: TNT-Cloning principle. One universal library in pSTART carries all DNA elements (Synthetic Biology Open Language, SBOL compliant) to produce multi-gene constructs by alternating use of two separate families of vectors, alpha (α; purple) and omega (Ω; orange) through the assembling loop.
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Enzymatic assembly of DNA molecules up to several hundred kilobases
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Iterative plug-and-play methodology for constructing and modifying synthetic gene networks
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Interactive assembly algorithms for molecular cloning
journal, April 2014
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Synthetic biology as it relates to CAM photosynthesis: challenges and opportunities
journal, February 2014
- DePaoli, Henrique C.; Borland, Anne M.; Tuskan, Gerald A.
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Type IIS Restriction Enzyme Footprinting I. Measurement of a Triple Helix Dissociation Constant With Eco57I at 25°C
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- Nucleic Acids Research, Vol. 24, Issue 12, p. 2435-2440
Synthetic gene expression perturbation systems with rapid, tunable, single-gene specificity in yeast
journal, December 2012
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- Nucleic Acids Research, Vol. 41, Issue 4
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REBASE—a database for DNA restriction and modification: enzymes, genes and genomes
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Arabidopsis Tetraspanins Are Confined to Discrete Expression Domains and Cell Types in Reproductive Tissues and Form Homo- and Heterodimers When Expressed in Yeast
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- Boavida, L. C.; Qin, P.; Broz, M.
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Self‐processing of ribozyme‐flanked RNAs into guide RNAs in vitro and in vivo for CRISPR‐mediated genome editing
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Standards for plant synthetic biology: a common syntax for exchange of DNA parts
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- Patron, Nicola J.; Orzaez, Diego; Marillonnet, Sylvestre
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- Sawers, Ruairidh J. H.; Farmer, Phyllis R.; Moffett, Peter
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A One Pot, One Step, Precision Cloning Method with High Throughput Capability
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A Modular Cloning System for Standardized Assembly of Multigene Constructs
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High Cleavage Efficiency of a 2A Peptide Derived from Porcine Teschovirus-1 in Human Cell Lines, Zebrafish and Mice
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