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Rapid genome recoding by iterative recombineering of synthetic DNA
Yu Heng Lau, Finn Stirling, James Kuo, Michiel A.P. Karrenbelt, Yujia A. Chan, Adam Riesselman, Connor A. Horton, Elena Schaefer, David Lips, Matthew T. Weinstock, Daniel G. Gibson, Jeffrey C. Way, Pamela A. Silver
doi: https://doi.org/10.1101/115493
Yu Heng Lau
aWyss Institute for Biologically Inspired Engineering, Harvard University, 3 Blackfan Circle, 5th Floor, Boston, MA 02115, USA
bDepartment of Systems Biology, Harvard Medical School, 200 Longwood Avenue, Alpert 536, Boston, MA 02115, USA
Finn Stirling
aWyss Institute for Biologically Inspired Engineering, Harvard University, 3 Blackfan Circle, 5th Floor, Boston, MA 02115, USA
bDepartment of Systems Biology, Harvard Medical School, 200 Longwood Avenue, Alpert 536, Boston, MA 02115, USA
James Kuo
aWyss Institute for Biologically Inspired Engineering, Harvard University, 3 Blackfan Circle, 5th Floor, Boston, MA 02115, USA
bDepartment of Systems Biology, Harvard Medical School, 200 Longwood Avenue, Alpert 536, Boston, MA 02115, USA
Michiel A.P. Karrenbelt
aWyss Institute for Biologically Inspired Engineering, Harvard University, 3 Blackfan Circle, 5th Floor, Boston, MA 02115, USA
cSystems and Synthetic Biology, Wageningen University, Dreijenplein 10, 6703 HB Wageningen, The Netherlands
Yujia A. Chan
aWyss Institute for Biologically Inspired Engineering, Harvard University, 3 Blackfan Circle, 5th Floor, Boston, MA 02115, USA
bDepartment of Systems Biology, Harvard Medical School, 200 Longwood Avenue, Alpert 536, Boston, MA 02115, USA
Adam Riesselman
dProgram in Biomedical Informatics, Harvard Medical School, Boston, MA 02115, USA
Connor A. Horton
aWyss Institute for Biologically Inspired Engineering, Harvard University, 3 Blackfan Circle, 5th Floor, Boston, MA 02115, USA
bDepartment of Systems Biology, Harvard Medical School, 200 Longwood Avenue, Alpert 536, Boston, MA 02115, USA
Elena Schaefer
aWyss Institute for Biologically Inspired Engineering, Harvard University, 3 Blackfan Circle, 5th Floor, Boston, MA 02115, USA
bDepartment of Systems Biology, Harvard Medical School, 200 Longwood Avenue, Alpert 536, Boston, MA 02115, USA
David Lips
aWyss Institute for Biologically Inspired Engineering, Harvard University, 3 Blackfan Circle, 5th Floor, Boston, MA 02115, USA
bDepartment of Systems Biology, Harvard Medical School, 200 Longwood Avenue, Alpert 536, Boston, MA 02115, USA
Matthew T. Weinstock
eSynthetic Genomics, Inc., 11149 North Torrey Pines Road, La Jolla, CA 92037, USA
Daniel G. Gibson
eSynthetic Genomics, Inc., 11149 North Torrey Pines Road, La Jolla, CA 92037, USA
fSynthetic Biology and Bioenergy Group, J. Craig Venter Institute, 4120 Capricorn Lane, La Jolla, CA 92037, USA
Jeffrey C. Way
aWyss Institute for Biologically Inspired Engineering, Harvard University, 3 Blackfan Circle, 5th Floor, Boston, MA 02115, USA
bDepartment of Systems Biology, Harvard Medical School, 200 Longwood Avenue, Alpert 536, Boston, MA 02115, USA
Pamela A. Silver
aWyss Institute for Biologically Inspired Engineering, Harvard University, 3 Blackfan Circle, 5th Floor, Boston, MA 02115, USA
bDepartment of Systems Biology, Harvard Medical School, 200 Longwood Avenue, Alpert 536, Boston, MA 02115, USA
Abstract
Genome recoding will provide a deeper understanding of genetics and transform biotechnology. We bypass the reliance of previous genome recoding methods on site-specific enzymes and demonstrate a rapid recombineering based strategy for writing genomes by Stepwise Integration of Rolling Circle Amplified Segments (SIRCAS). We installed the largest number of codon substitutions in a single organism yet published, creating a strain of Salmonella typhimurium with 1557 leucine codon changes across 200 kb of the genome.
Copyright
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. All rights reserved. No reuse allowed without permission.
Posted March 09, 2017.
Rapid genome recoding by iterative recombineering of synthetic DNA
Yu Heng Lau, Finn Stirling, James Kuo, Michiel A.P. Karrenbelt, Yujia A. Chan, Adam Riesselman, Connor A. Horton, Elena Schaefer, David Lips, Matthew T. Weinstock, Daniel G. Gibson, Jeffrey C. Way, Pamela A. Silver
bioRxiv 115493; doi: https://doi.org/10.1101/115493
Rapid genome recoding by iterative recombineering of synthetic DNA
Yu Heng Lau, Finn Stirling, James Kuo, Michiel A.P. Karrenbelt, Yujia A. Chan, Adam Riesselman, Connor A. Horton, Elena Schaefer, David Lips, Matthew T. Weinstock, Daniel G. Gibson, Jeffrey C. Way, Pamela A. Silver
bioRxiv 115493; doi: https://doi.org/10.1101/115493
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